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New Researchers Resource Center

Promega offers a wide range of resources dedicated to new researchers. We want to provide real support to help you from day one in the laboratory and all throughout your research. On this page, you will find the answers to the most common questions, access to tools, equipment, training and more! This page is designed to help:

  • • Master's candidates
  • • Doctoral Candidates (PhDs or pre-PhDs)
  • • Post-doctoral fellows
  • • Laboratory managers
  • • Research technicians

Promega is continuously creating new resources to help life science researchers. Be sure to connect with us so we can better support you in the long run!

  • Tip 1:

    How do you choose the best luciferase vector for your experiment?

    To help you, Promega has developed a tool called the "luciferase reporter vector selector" You will be able to select your vector by choosing the type of promoter, the application that interests you, the selection marker, or the type of luciferase.

    Customer and Technical Support

    Tip 2:

    What is multiplexing?

    Multiplexing different assays provides a set of data from the same sample. The main advantage is to have a better understanding of the event you are measuring in the context of another parameter, thus minimizing misinterpretation. The second advantage is that it allows you to standardize your results. To multiplex several assays, it is necessary that the signals of the various assays are spectrally or temporally distinct, the chemistries must be compatible, and be usable on the same well while being easily separated.

    Customer and Technical Support

    Tip 3:

    How do I authenticate my cell line?

    Cell line authentication need not be daunting: identification can be achieved by genetic profiling using polymorphic STRs (shorts tandem repeats). ATCC issued standards that recommend the use of at least 8 STRs loci for identification. The GenePrint10 kit contains 10 loci (TH01, TPOX, vWA, Amelogenin, CSF1PO, D16S539, D7S820, D13S317, D5S818 and D21S11) and has the power to confirm whether your cell line is the one you think. This kit can be used on all sequencers Applied Biosystems.

    Customer and Technical Support

    Tip 4:

    What are common issues that could hurt my luminescent assay results?

    Instruments with poor sensitivity may cause you to miss low-level samples that could be the “hit” you are looking for. Instruments with a narrow detection range limit the accuracy or reproducibility you needed to repeat your work. Finally, instruments that let the signal from bright wells spill into adjacent wells allow crosstalk to occur and skew experimental results, costing you time and leading to failed or repeated experiments.

    Customer and Technical Support

    Tip 5:

    How do I measure viability on 3D cells?

    CellTiter-Glo 3D is a bioluminescent assay that measures the level of ATP as an indicator of viability, thus making it possible to quantify cell viability within 3D spheroids. CellTiter-Glo 3D works with a single reagent in liquid form and generates a luminescent readout that is much more sensitive than colorimetric or fluorescence-based methods. The assay reagent penetrates large spheroids and has increased lytic capacity—allowing more accurate determination of viability compared to other assay methods.

    Customer and Technical Support

    Tip 6:

    How do you quickly and efficiently measure your level of ROS?

    ROS - "reactive oxygen species" produced by cells are molecule regulators involved in cellular signaling pathways. An excess of ROS can also induce cell death. The H₂O₂ level in a cell can reflect the general level of ROS in a cell. In the ROS-Glo H₂O₂ assay™ kit, A derivatized luciferin substrate is incubated with sample and reacts directly with H₂O₂ to generate a luciferin precursor. Addition of ROS-Glo™ Detection Solution converts the precursor to luciferin and provides Ultra-Glo™ Recombinant Luciferase to produce light signal that is proportional to the level of H₂O₂ present in the sample. With these assays, it is important to include multiple controls such as culture alone (which can contain or convert H₂O₂), with cells and a control positive as cells treated with menadione or pyrogallol.

    Customer and Technical Support

    Tip 7:

    Which plates do you use use for your fluorescence and luminescence measurements?

    In experiments involving fluorescence and luminescence measurements, a question may arise about which plate will allow for the best signal with low background noise. For luminescence, we recommend white opaque plates and for fluorescence measurements, we recommend opaque plates black. Using white plates to measure fluorescence will reduce the signal to background ratio but will give usable results. While using black plates for luminescence measurements will imply a decrease in the background noise but strong signal reduction (15 to 20 fold difference; white vs black). So, for experiments combining both a measure of fluorescence and luminescence, we recommend the use of white plates.

    Customer and Technical Support

    Tip 8:

    What additional factors should I consider when choosing a cell viability assay?

    Among the many factors to consider when choosing a cell-based assay, the primary concern for many researchers is the ease of use. When choosing an assay, the time required for reagent preparation and the total length of time necessary to develop a signal from the assay chemistry should be considered. Other factors to consider include sensitivity of detection, reagent stability and more. Check out the link below to learn more!

    Customer and Technical Support

    Tip 9:

    What is BRET?

    BRET stands for Bioluminescence Resonance Energy Tranfer. This technique measures an interaction between two proteins by donor energy transfer bioluminescent fluorescent acceptor when the two proteins interact. BRET allows a co-location in the cell of a PPI interaction.

    Customer and Technical Support

    Tip 10:

    How do I develop a test to study protein-protein interactions with NanoBRET?

    Promega scientist, Danette Daniels, explains how to set up a NanoBRET™ experiment, letting you fully explore protein interactions inside cells. Just click on the link below

    Customer and Technical Support
  • How to Interrogate Epigenetic Events in Live Cells

    Original Webinar Date: Tuesday, January 10, 2017

    Rather than alteration of the genetic code, epigenetics, or modifications of gene expression, is an interesting way that organisms develop a phenotype.

    Daniels.Danette

    Danette L Daniels, PhD

    Sr Research Scientist

    Understanding luminescent reporter assay design

    Original Webinar Date: Tuesday, March 8, 2016

    Luminescent reporter assays are powerful research tools for a variety of applications. This webinar explains critical aspects of reporter assay design to help you implement a robust assay that delivers meaningful results.

    Strayer.Carl

    Carl Strayer, PhD

    Lead Technical Services Scientist

    Factors to Consider for Designing and Optimizing Assays Applied to 3D Cultures

    Original Webinar Date: Tuesday, March 14, 2017

    3D cell culture models present a challenge for using assay chemistries and imaging methods that were originally designed for measuring events from 2D monolayers of cells. In this webinar, we will present factors to consider when designing and optimizing assays applied to 3D cell culture models, case studies.

    35323774-TerryRiss-600x750

    Terry Riss, PhD

    Global Strategic Marketing Manager, Cell Health

    Understanding the Role of the Inflammasome in Inflammation

    Original Webinar Date: Tuesday, January 12, 2016

    This webinar will discuss two important outcomes of inflammasome induction, caspase-1 activation and pyroptosis, and will present a new method for directly measuring caspase-1 activation.

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    Martha O'Brien, PhD

    Sr. Research Scientist

    How to measure live and dead cells using a plate reader

    Original Webinar Date: Tuesday, June 9, 2015

    This webinar will discuss how to use new assay methods that enable repeated "real time" measurement of live and dead cells using a standard plate reader while taking advantage of multiplexing opportunities. For more information, read the full abstract below.

    35323774-TerryRiss-600x750

    Terry Riss, PhD

    Global Strategic Marketing Manager, Cell Health

    Luciferase reporter assays tips and techniques

    Original Webinar Date: Tuesday, October 11, 2011

    Luciferase reporters are powerful and versatile tools for studying gene expression, signal transduction, and other cellular processes. Reporter assays can be complicated, but with the right knowledge and preparation researchers can apply these tools to produce a robust assay, and get the answer they need, the first time out.

    Strayer.Carl

    Carl Strayer, PhD

    Lead Technical Services Scientist

    Cell based assays to detect the mechanism of toxicity

    Original Webinar Date: Tuesday, March 13, 2012

    Investigating cytotoxicity in vitro often begins with measuring of the number of viable cells remaining in cultures incubated with test compounds for 2-3 days. Alternatively, the number of dead cells can be measured by detecting markers that leak from dead cells into the culture medium. Currently available assay technologies can go beyond providing simple “live or dead” information to indicate whether cells have died by apoptosis or necrosis.

    35323774-TerryRiss-600x750

    Terry Riss, PhD

    Global Strategic Marketing Manager, Cell Health

  • Designing a bioluminescent reporter assay

    When designing a bioluminescent reporter assay, normalization should be an essential part of the assay plan in order to control for variables other than those being tested. Here we show how to normalize your bioluminescent reporter assay experiments.

    Finding a Connection Between Glucose Metabolism and Macrophage Activation

    A decade ago, immunology and metabolism were seen as two distinct areas of study. However, we now know that specific metabolic activities are required for proper immune cell differentiation and function. In tumor microenvironments, immune cells may even alter their metabolism to compete with tumor cells for limiting nutrients.

    Revealing time of death ; the microbiome edition

    Forensic analysts have long sought precision when determining time of death. While on crime scene investigation television shows, the presence of insects always seems to reveal when a person died, there are many elements to account for, and the probable date may still not be accurate.

    Don’t let 3 common issues hurt your luminescent assay results

    There is a lot riding on your luminescent assay results. Each plate represents precious time, effort and resources. Did you know that there are three things about your detection instrument that can impact how much useful information you get from each plate?

    The randomness of cancer

    A major scientific study grabbed headlines recently, and the implications of its findings may affect many of us, if not all of us. In a paper published in Science by Cristian Tomasetti, Lu Li and Bert Vogelstein of Johns Hopkins University, the authors report that nearly two-thirds of known cancer causing mutations can be attributed to random mistakes that occur during DNA replication.

    The Cell Line Identity Crisis: Old problems, new concerns

    If you work with cell lines you may have paid attention to the dramatic headline published last month in the online journal STAT, Thousands of studies used the wrong cells, and journals are doing 29981164-Whistlenothing.” In their column The Watchdogs.

    Using the NanoBiT™ System to Study Protein Interaction Dynamics In Live Cells

    NanoLuc® Binary Technology, or NanoBiT, is a new binary complementation system based on NanoLuc® Luciferase (1). NanoLuc® Luciferase is a 19 kDa enzyme that produces very bright, glow-type luminescence. To create a complementation system, We separated NanoLuc® Luciferase into two subunits, a small peptide and a larger protein domain.

  • How to choose the best luciferase vector for your experiment?

    You will be able to select your vector by choosing the type of promoter, the application that interests you (eg imaging), the selection marker or the type of luciferase

  • Training

    NanoBiT ™: a new technology to study protein / protein interactions

    With NanoBIT ™ (NanoLuc® Binary Technology) Promega offers a new solution for researchers to follow the dynamics of protein / protein interactions within cells. Explore protein interactions by benefiting from a unique bioluminescence-based complementation system for your applications: receptor dimerization, receptor/ligand interaction, signaling event tracking.

  • Tip 1:

    Can PCR products generated with GoTaq® G2 DNA polymerase be used to make TA cloning?

    Yes. GoTaq® DNA Polymerase is a robust formulation of unmodified Taq Polymerase. GoTaq®DNA Polymerase lacks 3’ →5’ exonuclease activity (proof reading) and also displays non-template–dependent terminal transferase activity that adds a 3′ deoxyadenosine (dA) to product ends. As a result, PCR products amplified using GoTaq® DNA Polymerase will contain A-overhangs which makes it suitable for T-vector cloning. We have successfully cloned PCR products generated using GoTaq® and GoTaq® Flexi DNA Polymerases into the pGEM®-T (Cat.# A3600), pGEM®-T Easy (Cat.# A1360) and pTARGET™ (Cat.# A1410) Vectors.

    Customer and Technical Support

    Tip 2:

    What is the easiest way to clone PCR Products?

    The simplest way to clone PCR Products is to amplify the product using thermostable polymerases such as Taq, Tfl or Tth polymerase. These polymerases add a single deoxyadenosine to the 3´-end of the amplified products (3´-end overhang), and can be cloned directly into a linearized T-vector. Click on the link below to get answers to our most common questions!

    Customer and Technical Support

    Tip 3:

    What are some considerations to make to improve my qPCR results?

    There are a number of parameters that can influence the success of your qPCR assay. For one, sample purity is crucial! Use precautions to minimize the potential for sample cross-contamination. Some practices to help with contamination problems include using aerosol resistant tips, wearing gloves and changing them frequently. Establish dedicated pre- and post-amplification areas so that you are never opening reaction tubes after amplification in the area where you will be setting up new reactions. Click the link below for the Top 10 Tips for improving your qPCR or RT-qPCR assays!

    Customer and Technical Support

    Tip 4:

    Do I need to add CXR reference dye in my GoTaq® qPCR reactions?

    GoTaq® qPCR Master Mix already contain a low level of carboxy-X-rhodamine (CXR); (identical to ROX) which makes it compatible with most qPCR instruments. If your thermocycler requires high concentrations of CXR as some Applied Biosystems instruments do, simply add the separate tube of CXR reference dye (included in kit). Do not hesitate to contact technical support or consult the technical manual to find out if your instrument requires a small or large amount of CXR.

    Customer and Technical Support

    Tip 5:

    Which competent bacteria can I use with pGEM®-T for the selection of positive clones?

    Use the high efficiency JM109 bacteria that comes with the kit, or any other bacterium that has in its genotype LacZΔM15 and allows blue/white screening on your petri dishes.

    Customer and Technical Support
  • Introduction to Real-Time PCR: Basic Principles and Chemistries

    Original Webinar Date: Tuesday, August 9, 2016

    Real-Time PCR (qPCR) is the workhorse of most gene expression analysis. This webinar will provide an overview of basic qPCR principles compared to traditional PCR and common terms used in assay design and analysis.

    Leta-Steffen

    Leta Steffen, PhD

    Applications Scientist Promega Corporation

    Guidelines for developing Robust and Reliable PCR assays

    Original Webinar Date: Tuesday, December 6, 2016

    A robust and reliably performing qPCR assay is dependent on an optimal balance of the components in the amplification reaction environment. Review how PCR components influence qPCR assay performance, Learn how to optimize a qPCR assay ...

    Leta-Steffen

    Leta Steffen, PhD

    Applications Scientist Promega Corporation

    To NanoDrop® or Not to NanoDrop®: Choosing the Most Appropriate Method for Nucleic Acid Quantitation

    Original Webinar Date: Tuesday, February 14, 2017

    Success or failure in DNA/RNA analysis applications often comes down to whether or not the appropriate amount of input nucleic acid is used, but nucleic acid quantitation methods reveal different information about a sample. This webinar will review absorbance, fluorescent nucleic acid-binding dyes and qPCR quantitation methods and present the advantages and ...

    2219_Wieczorek_Doug_BIG

    Doug Wieczorek, PhD

    Sr. Applications Scientist

    The Hows and Whys of Early Steps in RNA Analysis

    Original Webinar Date: Tuesday, August 11, 2015

    Learn about the early steps of purification, protection from degradation and quantitation to improve your downstream analysis.

    Leta-Steffen

    Leta Steffen, PhD

    Applications Scientist Promega Corporation

  • Is qPCR for You ?

    Why qPCR?

    If you need to detect DNA sequences of low abundance in a sample or quantify the amount of DNA or gene expression levels in a sample, qPCR can precisely measure the PCR products. This technique, which measures the accumulation of PCR product as it occurs during amplification, can deliver a great deal of information about a sample in a short amount of time.

    Real-Time qPCR : Considerations for comparing Reagent Performance

    Real-time quantitative PCR (qPCR) is a powerful tool to detect and quantify nucleic acids. By incorporating fluorescently labeled probes or fluorescent double-stranded DNA (dsDNA)-binding dyes into the PCR, product formation can be monitored following each PCR cycle. Promega GoTaq® qPCR Master Mixes for dye-based or probe-based product detection are optimized for fast and reproducible qPCR assays. In this guide, we outline some of the most important considerations.

    Real-Time PCR: Guidelines for a Comparison of Reagent Performance

    A protocol for testing previously optimized qPCR assays with a new qPCR master mix to look at several factors of reagent performance including assay specificity, reproducibility, linearity, sensitivity and efficiency.

    How Do I Choose the Right GoTaq® Product to Suit My Needs for EndPoint PCR?

    We offer a wide array of GoTaq® DNA Polymerases, Buffers and Master Mixes, so we frequently answer questions about which product would best suit a researcher’s needs. On the product web page, you can filter the products by clicking the categories on the left hand side of the page to narrow down your search. Here are some guidelines with the relevant categories in bold type to help you select the match that will best suit your PCR application.

  • Amplification Product Selector

    To find the appropriate amplification product for your experiment, filter by the radio button items on the left side. If you have a question about a particular product that is not included in this selector, please contact Technical Services to find the solution that is right for you.

  • Training

    Best practices to optimize your qPCR reactions

    This video training addresses the fundamental principles of real-time PCR, from its implementation to the interpretation of results.

  • Tip 1:

    What reference protein extract can I use to test protein mass spec sample preparation procedures and validate mass spec instrument performance?

    There are 2 types of protein extracts prepared from yeast (Saccharomyces cerevisiae) or human (K562) cells that can serve as reference material: One, is a crude protein extract from whole cells that validates your preparation protocol before the analysis. Another, is a protein extract predigested with Trypsin/Lys-C and treated by DTT and iodoacetamide. This validates your instrument and confirms the reproducibility of your results from one analysis to another. These extracts are freeze-dried, which allows them to be easily suspended in TFA or formic acid before injection into the device.

    Customer and Technical Support

    Tip 2:

    What TNT® system do I use for which protein size?

    Depending on the size of the proteins, several TNT® transcription/translation systems can be used:
    - For proteins smaller than 40 kDa, we suggest you use TNT® based on wheat germ extracts.
    - For proteins with an average size of 40 to 80 kDa, Promega’s TNT® systems based on wheat germ extracts or reticulocyte lysates are suggested.
    - For proteins larger than 80 kDa, we recommend the use of TNT® systems with reticulocyte lysates. To increase the amount of protein completely translated, it is also recommended to work at 25°C instead of 30°C.

    Customer and Technical Support

    Tip 3:

    Which affinity tag do I choose for my proteins?

    Many fusion "tags" have been developed for different applications, such as for the purification of proteins or the study of their intracellular localization. Promega’s new HaloTag® is fused to a protein and will bind covalently to the different types of ligands available: permeant or non-permeant fluorescent markers, magnetic beads, biotin, etc. So, with the same construct and interchanging only the ligand, you can use it for all kinds of applications.

    Customer and Technical Support

    Tip 4:

    How can I digest my proteins quickly and efficiently?

    Try Promega’s new Rapid Trypsin & Trypsin/Lys-C Kits! This new product is designed to shorten protein digestion times to 60 minutes versus the typical 4–18 hours. Protein digestion with these kits follows a simple-to-use protocol that is both fast and efficient. The protocol is flexible, can accommodate a large range of sample volumes and protein concentrations and requires no special laboratory equipment or off-line desalting.

    Customer and Technical Support

    Tip 5:

    What are the advantages of using a TNT® coupled transcription/translation system over standard systems?

    For most gene constructs, the TnT® Quick reaction produces significantly more protein (two- to sixfold) in a 60- to 90-minute reaction than a standard in vitro rabbit reticulocyte lysate reaction using RNA templates.

    Customer and Technical Support

    Tip 6:

    Are the post translational modifications of my protein ensured in TNT® in vitro transcription / translation systems?

    Yes, most modifications (phosphorylation, acetylation, ubiquitination, etc.) are provided by rabbit reticulocyte lysate or wheat germ extract. The addition of microsomal membranes with reticulocyte lysate allows modifications such as N-glycosylation or signal peptide cleavage.

    Customer and Technical Support
  • Tools for improved protein mass spec sample preparations

    Original Webinar Date: Tuesday, May 24, 2016

    This webinar will discuss common shortcomings of protein mass spec sample preparation and provide solutions to these shortcomings. Common shortcomings of protein mass spec sample preparation include incomplete digestion, lengthy and laborious procedure, low protein sequence coverage, inefficient peptide recovery ...

    1938_Saveliev_Sergei_BIG

    Sergei Saveliev, PhD

    Sr. Research Scientist

    Beyond Trypsin: Strategies to Improve Mass Spec Sequence Coverage and Post-Translation...

    Original Webinar Date: Tuesday, July 10, 2012

    Proteomics research largely focuses on using biological mass spectrometry (MS) to sequence individual proteins and whole proteomes. The target proteins are first digested with proteases, in which trypsin has historically been used due to its high activity, specificity, optimal size, and C-terminally charged peptides for MS analysis.

    3504_Rosenblatt_Mike_BIG

    Mike Rosenblatt, Ph.D.

    Overcoming challenges of protein analysis in mammalian systems

    Original Webinar Date: Tuesday, January 10, 2012

    The growing trend towards study of mammalian proteins in their native environment has presented specific challenges in the areas of protein expression and purification, monitoring trafficking events both spatially and temporally, and isolation of intact intracellular protein complexes. The HaloTag® technology, a multifunctional fusion protein tag, allows researchers to....

    Daniels.Danette

    Danette L Daniels, PhD

    Sr Research Scientist

    In Vitro and In Vivo Methods to Study Protein Protein Interactions

    Original Webinar Date: Tuesday, November 8, 2011

    Proteins rarely act alone. Interactions between proteins is an essential part of many biological functions. Deciphering and characterizing specific interactions is a common part of many laboratory investigations. This seminar will discuss cell-free translation systems and how rapid protein production can aid in these investigations through common applications like GST pulldowns and co-immunoprecipitation.

    3621_Brazas_Rob_BIG

    Dr. Rob Brazas

    In Vitro, Cell-Free Protein Expression - How It Helps Speed Up Your Research

    Original Webinar Date: Tuesday, January 14, 2014

    In vitro, cell-free protein expression is a convenient method for rapidly producing protein in 1-4 hours. This webinar will provide an overview of the various cell-free expression systems available and methods to label and detect the expressed proteins.

    49_Kobs_Gary_BIG

    Gary Kobs

    Proteomics Strategic Marketing Manager

  • Why wait? Sample preparation in as little as 30 minutes

    While many proteases are used in bottom-up mass spectrometric (MS) analysis, trypsin (4,5) is the de facto protease of choice for most applications. There are several reasons for this: ...

    Moving out of the cell : advantages of cell free expression

    Cell-free protein expression is a simplified and accelerated avenue for the transcription and/or translation of a specific protein in a quasi cell environment. An alternative to slower, more cumbersome cell-based methods, cell-free protein expression methods....

  • Mass Spectrometry Proteases Selector

    We offer a variety of proteases used in the characterization of proteins by mass spec. This selector will help you to select the optimal protease for your experimental studies.

  • Under Construction.

    We are currently updating this section, check back soon!

    En Construction
  • Under Construction.

    We are currently updating this section, check back soon!

    En Construction
  • Tip 1:

    What are the factors influencing the percentage of transfected cells?

    The efficiency of the transfection will depend on different parameters:
    - Cell type and cell health (appropriate culture medium, growth, serum etc.)
    - The confluence of the culture (in general, cells are used between 40 and 80% of confluence)
    - The number of passages (avoid too many passages <50)
    - The quality and quantity of DNA, RNA or siRNA (no protein contaminants, endotoxin, etc.)
    - Ratio DNA / transfection compound (1: 1 to 2: 1)
    - The transfection time (from 30 min to several days if stable)

    Customer and Technical Support

    Tip 2:

    How do I develop LigaFast ™ fast ligation?

    Ligation involves inserting a piece of DNA into a vector by binding the 5'-phosphate and 3'-hydroxyl ends of the adjacent nucleotides:
    - We recommend starting with a 1:2 molar ratio of vector:insert DNA when cloning a fragment into a plasmid vector
    - Click here to view remaining steps.

    Customer and Technical Support

    Tip 3:

    How can I optimize transfection efficiency?

    Due to the variety in cell types, each will have different transfection efficiencies (lineage or primary cells). It's important to:
    - Choose the best transfection method (chemical, liposomes, electroporation, etc.)
    - Ensure your nucleic acids are of good quality
    - Test different transfection agent/DNA ratios
    - Check the efficiency of transfection (second reporter, standardization etc.)

    Customer and Technical Support

    Tip 4:

    How does multiplexing allow for easy optimization?

    Keep optimization simple by using reporter and viability assays that can be measured in the same sample. The ONE-Glo™ Luciferase Assay System for measuring firefly luciferase (Cat.# E6110) and the CellTiter-Fluor™ Cell Viability Assay (Cat.# G6080) are examples of compatible assays. By multiplexing these two assays, both reporter activity and viability can be measured in the same well of a 96-well plate in less than an hour. No medium changes or washing is needed.

    Customer and Technical Support
  • Under Construction.

    We are currently updating this section, check back soon!

    En Construction
  • Improving the success of your transfection

    Not every lab has a tried and true transfection protocol that can be used by all lab members. Few researchers will use the same cell type and same construct to generate data. Many times, a scientist may need to transfect different constructs or even different molecules (e.g., short-interfering RNA [siRNA]) into the same cell line, or test a single construct in different cultured cell lines.

    General Considerations for Transfection

    Many studies, from reporter assays to protein localization to BRET and FRET, require successful transfection first. Yet, transfection can be tricky and difficult. There are many considerations when planning transfection of your cells including reagent selection, stable or transient experiment, type of molecule and endpoint assay used. Here we discuss these considerations to help you […]

    The Keys to Successful Transfection

    Transfection, like many techniques in the lab, can be an art, especially for stubborn cell lines. To maximize transfection efficiency, you must have the right conditions and, for some cell lines it seems, the right touch. If you’re lucky, you are working with less finicky cells that transfect well regardless of conditions. If you’re not […]

    Protocols for transfection and information on optimization

    Transfection is a method that neutralizes or obviates the issue of introducing negatively charged molecules (e.g., phosphate backbones of DNA and RNA) into cells with a negatively charged membrane. Chemicals like calcium phosphate and DEAE-dextran or cationic lipid-based reagents coat the DNA, neutralizing or even creating an overall positive charge to the molecule (Figure 12.1). This makes it easier for the DNA:transfection reagent complex to cross the membrane.

    Reverse Transfection Using FuGENE® 6 and FuGENE® HD

    In reverse transfection protocols, cells are added directly to a plate containing the transfection reagent:DNA mixture and then assayed on the second day. This approach reduces the experimental time by a day compared to standard transfection protocols. Reverse transfections are rapid, convenient and easily automated, and are frequently used for high-throughput experiments. We tested the ability of FuGENE® 6 and FuGENE® HD to “reverse” transfect three cell lines and ...

    Use the Best Cell Line for Your Study

    FuGENE® HD Transfection Reagent offers unrivaled transfection performance with minimal impact on cell physiology (Jacobson et al. [2009] Biotechniques 47, 617–24). It is ideal for use in a variety of challenging applications, including transfection of cancer model cell lines, insect cells, stem cells, and for virus and protein production.

    Optimize Transfection of Cultured Cells

    Protein expression from plasmid DNA transfected into cultured cells is a powerful tool to study protein function and monitor intracellular events. Optimal protein expression and cell viability require optimizing transfection conditions. We discuss how to optimize transfection conditions using FuGENE® HD Transfection Reagent and the factors that are most likely to influence success.

    From Napkin Sketch to “Custom Kit”: CloneWeaver® Workflow Builder Gets Your Cloning Organized

    Let’s face it, most lab techs and purchasing agents aren’t all that happy when you send them an Instagram picture of your latest lunchroom-napkin cloning strategy as your order form for your next big cloning experiment. So we have created the CloneWeaver® Workflow Builder. You can transfer your brilliance easily from that lunchroom napkin to […]

  • Your Transfection Assistant

    The Transfection Assistant helps you optimize transfection protocols using information from the published literature. It provides information on transfection of specific cell types, including the amount of nucleic acid and ratio of reagent:nucleic acid used for specific plate formats, and provides links to the primary literature.

    Select a cell line and transfection reagent to search the database or select “View All” to see the entire database contents.

    FuGENE® HD Protocol Database

    The FuGENE® HD Protocol Database is a collection of protocols that guide the transfection process for a variety of cell lines. These protocols were developed by Promega Corporation or Fugent, L.L.C., and the outlined conditions gave appreciable transfection results and little toxicity. Further optimization may be possible if transfection efficiency or viability needs to be maximized. Most protocols were tested in a 96-well plate format; therefore, protocols generated for other plate types are based on calculations and are suggested starting points to achieve good results.

    CloneWeaver® Workflow Builder

    Powering Your Cloning Workflow

    Product selection has never been easier

    Molecular Cloning is the process of producing recombinant DNA and transforming into host organisms to replicate and make more copies. Every cloning project is unique. Once you have designed the cloning scheme, gathering all of the required reagents to get you from construct to expression and analysis is not trivial. CloneWeaver helps you build a customized cloning "kit" with all of the items your cloning scheme requires. Purchase your selection instantly, save it or email it to a purchasing agent. Soon you'll have everything you require to create the construct you need.

  • Training

    Introduction to transfection

    Transfection is a method of introducing nucleic acids into eukaryotic cells by minimizing undesirable effects such as cytotoxicity, modification of gene expression or cell death.

Additional Resources

Stretch your Startup Budget

Preparing a lab can challenge any researcher. We’re committed to making it easier by helping you stretch your budget with our New Lab Startup Program. Whether you’re a part of your lab’s research, procurement, or administration team you are eligible to participate in the New Lab Startup Program. When you register for our program, you’ll work with a Scientific Specialist on product recommendations based on your unique research needs. Let us help you focus on the work instead of worrying about stocking reagents. Register for the this easy program to get started!

Promega Training Support Program

If you are an instructor for a course using DNA, RNA, protein or cell-based techniques, you may qualify for assistance available through the Promega Training Support Program. The program offers a 50% discount off list price up to a maximum of $2,000.00 in total discounts to make your training budget go farther. See terms and conditions.

Helix® On-site Stocking

With Helix® freezers, refrigerators and cabinets, you get reliable 24/7 access to the products you need when and where you need them. Reduce your paperwork, save time and money and avoid delays to get back to your research quickly.

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