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CloneWeaver® Workflow Purchasing Tool

Powering Your Cloning Workflow

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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. Read More

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.

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PCR Cloning

Name Description Part Number
Set of dATP, dCTP, dGTP, dUTP Open/Close Add
Highly pure (≥99% triphosphate)
Each dNTP supplied at 100mM in water for ease-of-use
Incorporate dUTP into PCR products and treat with uracil-N-glycosylase to inhibit subsequent amplifications

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5. dUTP (2'-Deoxyuridine, 5'-Triphosphate) can be used in place of dTTP in PCR and RT-PCR protocols to prevent carryover from previous amplifications. The substitution of dUTP for dTTP in PCR results in uracil-containing PCR products that are suitable for most standard applications. The enzyme uracil-N-glycosylase (UNG, also referred to as UDG) can be added to a PCR premix to excise uracil from any contaminating PCR product, thereby preventing false positives. Each lot of dUTP is function-tested to ensure specific DNA amplification and the absence of nuclease activity.

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Set of dATP, dCTP, dGTP, dUTP Open/Close Add
Highly pure (≥99% triphosphate)
Each dNTP supplied at 100mM in water for ease-of-use
Incorporate dUTP into PCR products and treat with uracil-N-glycosylase to inhibit subsequent amplifications

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5. dUTP (2'-Deoxyuridine, 5'-Triphosphate) can be used in place of dTTP in PCR and RT-PCR protocols to prevent carryover from previous amplifications. The substitution of dUTP for dTTP in PCR results in uracil-containing PCR products that are suitable for most standard applications. The enzyme uracil-N-glycosylase (UNG, also referred to as UDG) can be added to a PCR premix to excise uracil from any contaminating PCR product, thereby preventing false positives. Each lot of dUTP is function-tested to ensure specific DNA amplification and the absence of nuclease activity.

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Set of dATP, dCTP, dGTP, dTTP Open/Close Add
Provided at 100mM in water at pH 7.5

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5.

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RNasin Ribonuclease Inhibitors Open/Close Add
Native, recombinant and oxidation-resistant forms available
Does not inhibit SP6, T7 or T3 RNA Polymerase; GoScript™ Reverse Transcriptase, AMV or M-MLV Reverse Transcriptase; or Taq DNA polymerase.
Use in many downstream assays; inhibits a broad spectrum of eukaryotic RNases over a wide pH range (pH 5–8)
Maintains inhibitory activity over a wide temperature range

RNases are ubiquitous and can cause RNA degradation and compromise RNA integrity. Native and Recombinant RNasin Inhibitors are 50kDa proteins that inhibit RNase A family and human placental RNases by noncovalently binding to RNases in a 1:1 ratio. For downstream applications such as GoScript Reverse Transcriptase, AMV/M-MLV reverse transcriptases, SP6, T7/T3 RNA polymerase, and Taq DNA polymerases, Recombinant RNasin Inhibitor does not inhibit RNase T1, S1 nuclease, RNase from Aspergillus, RNase H, RNase ONE Ribonuclease and enzymes. RNasin Plus RNase Inhibitor is a recombinant mammalian RNase inhibitor that is expressed as a soluble protein in E. coli, allowing easy purification through a combination of ion exchange and hydrophobic interaction chromatography. The protein is capable of inhibiting eukaryotic RNases (e.g., RNase A and RNase B) similarly to human placental RNase inhibitor. RNasin Plus RNase Inhibitor is tested in RT-PCR and compatible with enzymes such as AMV, M-MLV and ImProm-II Reverse Transcriptases or Taq and Tfl DNA Polymerases. RNasin Plus RNase Inhibitor also is tested and compatible with quantitative, real-time RT-PCR in a TaqMan assay. RNasin Plus RNase Inhibitor offers increased resistance to oxidation over the human version of the protein. Two cysteines in the human protein have been identified as especially sensitive to oxidation and react by forming a disulfide bond that can block the active site of the inhibitor. RNasin Plus, through natural amino acid diversity, lacks the ability to form this site-blocking disulfide. In addition, the new protein has characteristics never before realized, including continued inhibition of RNases above 50°C. Heating solutions of RNasin Plus and RNase followed by cooling does not result in the reappearance of RNase activity—even when the solution is heated above the denaturation temperature of the RNasin Plus protein alone. This allows RNasin Plus to protect RNA species prior to, during and after heating, even at temperatures normally used during first-strand DNA synthesis in RT-PCR. Solutions heated up to 70°C for 15 minutes did not result in RNase reactivation.

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Reverse Transcription System Open/Close Add

The Reverse Transcription System provides reagents to efficiently reverse transcribe RNA into cDNA in 15 minutes. The cDNA prepared from each reaction using this system may be used directly in multiple PCR amplifications using Taq DNA polymerase. The AMV Reverse Transcriptase synthesizes single-stranded cDNA from total or poly(A)+ RNA. Both Oligo(dT)15 and Random Primers are included, allowing cDNA synthesis from virtually any RNA source. The system contains sufficient reagents for 100 cDNA synthesis reactions, processing 1ug of RNA per reaction. Each cDNA synthesis reaction may be divided and used in up to 20 separate PCR amplifications. A polyadenylated 1.2kb RNA transcript is provided as a control template for cDNA synthesis.

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M-MLV Reverse Transcriptase, RNase H Minus, Point Mutant Open/Close Add

Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus (M-MLV RT [H-]), Point Mutant, is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long RNA templates (>5kb). The lack of RNase H activity is beneficial for this application, as RNase H can start to degrade templates when incubation times are long, as they may be when synthesizing long cDNAs. Although many researchers are successful in using M-MLV RT (H+) for analytical and some preparative cDNA applications, reverse transcriptases lacking RNase H activity provide another option to prepare long cDNAs and libraries containing a high percentage of full-length cDNA.

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M-MLV Reverse Transcriptase, RNase H Minus Open/Close Add
Supplied at a concentration of 100–200u/μl

Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus (M-MLV RT [H-]), is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long messenger RNA templates (>5kb). This form of M-MLV Reverse Transcriptase has been genetically altered to remove the associated RNase H activity. Although many researchers are successful in using M-MLV RT (H+) for analytical and some preparative cDNA applications, reverse transcriptases lacking RNase H activity provide another option to prepare long cDNAs and libraries containing a high percentage of full-length cDNA.

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M-MLV Reverse Transcriptase Open/Close Add
Enzyme supplied at a concentration of 200u/μl

Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long messenger RNA templates (>5kb). The enzyme is a product of the pol gene of M-MLV and consists of a single subunit with a molecular weight of 71kDa. The RNase H activity of M-MLV RT is weaker than that of the commonly used Avian Myeloblastosis Virus (AMV) reverse transcriptase.

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ImProm-II Reverse Transcription System Open/Close Add

The ImProm-II Reverse Transcription System produces efficient, robust synthesis of first-strand cDNA in preparation for PCR amplification. The components of the ImProm-II Reverse Transcription System can be used to reverse transcribe RNA templates starting with total RNA, poly(A)+ mRNA or synthetic transcript RNA. The optimized reaction buffer and powerful ImProm-II Reverse Transcriptase provided in the ImProm-II System together enable robust, full-length cDNA synthesis for the reproducible analysis of rare or long messages. The cDNA synthesis conditions were formulated for standalone applications or for easy transition to gene-specific target amplification. An aliquot of the reverse transcription reaction (1-20ul) can be amplified directly using Taq DNA polymerase in coupled or uncoupled PCR.

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GoTaq Reaction Buffers Open/Close Add
Available with green dye for direct-to-gel convenience, or in colorless format for applications where absorbance or fluorescence measurements are necessary prior to PCR cleanup
Contain 7.5mM MgCl2 for a final concentration of 1.5mM in a 1X reaction

The 5X Green GoTaq Reaction Buffer contains two dyes (a blue dye and a yellow dye) that separate during electrophoresis to show migration progress. The buffer also contains a compound that increases sample density. This means that samples can be loaded directly onto gels without the need for loading dye. The blue dye migrates at the same rate as a 3-5kb DNA fragment in a 1% agarose gel. The yellow dye migrates at a rate faster than primers (<50bp) in a 1% agarose gel. The 5X Colorless GoTaq Reaction Buffer has the same formulation as the 5X Green GoTaq Reaction Buffer but does not contain dyes and is recommended for any applications where absorbance or fluorescence measurements are necessary prior to PCR cleanup. Both buffers are supplied at pH 8.5.

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GoTaq Long PCR Master Mix Open/Close Add
Optimized blend of thermostable DNA polymerases with enhanced processivity and proofreading
Hot-start master mix for convenient room temperature setup
Optimized components for enhanced yield, sensitivity and specificity

GoTaq Long PCR Master Mix contains the high-performance GoTaq Hot Start Polymerase in a specially formulated mixture with a proprietary thermostable proofreading polymerase. This optimized enzyme mixture allows efficient amplification of up to 40kb from lambda DNA or 30kb from human genomic DNA. The presence of a proofreading enzyme to repair DNA mismatches and a highly processive polymerase allows the polymerase to continue to elongate the DNA much further, resulting in longer DNA amplification. The optimized formulation of the GoTaq Long PCR Master Mix components enables simple reaction setup and provides consistently efficient, accurate and robust amplification of long DNA amplicons.

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GoTaq Hot Start Polymerase Open/Close Add
Hot Start Taq minimizes nonspecific amplification and primer-dimers
Optimize reaction conditions with supplied Flexi Reaction Buffer and MgCl2 solution
Direct-to-gel convenience
Supplied at a concentration of 5u/µl

GoTaq Hot Start Polymerase contains the high-performance GoTaq DNA Polymerase bound to a proprietary antibody that blocks polymerase activity. The polymerase activity is restored during the initial denaturation step when the amplification reactions are heated at 94-95 degrees C for two minutes. This enables hot-start PCR, where polymerase activity is eliminated or minimized at temperatures below 70 degrees C. GoTaq Hot Start Polymerase exhibits 5' --> 3' exonuclease activity. The enzyme is supplied with a tube of 25mM MgCl2 to optimize the magnesium concentration in your reactions. It is also supplied with 5X Green GoTaq Flexi Buffer and 5X Colorless GoTaq Flexi Buffer. The buffers contain a compound that increases sample density so that samples sink easily into wells of an agarose gel. The green buffer also contains two dyes (yellow and blue) that separate to allow easy monitoring during electrophoresis. Use the green reaction buffer for direct-to-gel analysis after amplification and the colorless reaction buffer for amplifications where the dyes may interfere with post-amplification analysis such as fluorescence or absorbance testing.

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GoTaq Hot Start Master Mixes Open/Close Add
All-in-one master mix; just add template and primers
Minimize nonspecific amplification and primer-dimers
Use the green master mix for direct-to-gel analysis, and use the colorless master mix when post-amplification fluorescence or absorbance analysis is required

GoTaq Hot Start Master Mixes are premixed, ready-to-use solutions containing GoTaq Hot Start Polymerase, magnesium, dNTPs and buffer. Reactions can be set up in less than a minute at room temperature; simply add your template, water and primers. GoTaq Hot Start Hot Start Polymerase contains the high-performance GoTaq DNA Polymerase bound to a proprietary antibody that blocks polymerase activity. The polymerase activity is restored during the initial denaturation step when the amplification reactions are heated at 94-95 degrees C for two minutes. This enables hot-start PCR, where polymerase activity is eliminated or minimized at temperatures below 70 degrees C. GoTaq Hot Start Polymerase exhibits 5' --> 3' exonuclease activity. The master mixes are available in green or colorless formats. The green master mix contain a compound that increases sample density so that samples sink easily into wells of an agarose gel as well as two dyes (yellow and blue) that separate to allow easy monitoring during electrophoresis. Use the green master mix for direct-to-gel analysis after amplification and the colorless master mix for amplifications where the dyes may interfere with post-amplification analysis such as fluorescence or absorbance testing. GoTaq Hot Start Master Mixes offer the specificity and sensitivity of an antibody-based hot-start polymerase in a convenient, easy-to-use, time-saving format.

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GoTaq G2 Hot Start Polymerase Open/Close Add
Hot-start Taq eliminates nonspecific PCR amplification
Detects low-copy-number targets
Allows optimization of reactions with Flexi Reaction Buffer and separate MgCl2
Supplied at a concentration of 5u/μl

For superior convenience and improved yield, sensitivity and specificity, choose GoTaq G2 Hot Start Polymerase, which is bound to a proprietary antibody that blocks activity. Activity is restored during initial denaturation, but is inhibited below 70 degrees C, allowing room-temperature reaction setup. GoTaq G2 Hot Start Polymerase is supplied with 5X Green GoTaq Flexi Buffer, 5X Colorless GoTaq Flexi Buffer and 25mM MgCl2. Hot-start PCR is advantageous for some amplification targets because it may eliminate or minimize primer-dimer and nonspecific products. In some cases, hot-start PCR may improve yields. GoTaq G2 Hot Start Polymerase exhibits 5' --> 3' exonuclease activity.

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GoTaq G2 Hot Start Master Mixes Open/Close Add
All-in-one master mix; just add template and primers
Eliminate nonspecific amplification with a hot start enzyme
Use the green master mix for direct-to-gel analysis, and the colorless master mix when post-amplification fluorescence or absorbance analysis is required

GoTaq G2 Hot Start Master Mixes are ready-to-use mixes containing GoTaq G2 Hot Start Polymerase, buffer, dNTPs and optimized magnesium—you only need to add primers and template and go! The GoTaq G2 Hot Start Green Master Mix also contains a gel loading dye to facilitate downstream gel analysis. The GoTaq G2 Hot Start Colorless Master Mix contains no gel loading dye and is used when downstream applications require fluorescence or absorbance readings without purification. GoTaq G2 DNA Polymerase is bound to a proprietary antibody that blocks polymerase activity until the initial denaturation step of the amliication reaction, allowing hot-start PCR. Hot-start PCR is advantageous for some amplification targets because it may eliminate or minimize primer-dimer and nonspecific products. GoTaq G2 Hot Start Polymerase exhibits 5' --> 3' exonuclease activity.

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GoTaq G2 Flexi DNA Polymerase Open/Close Add
Full-length Taq DNA polymerase that exhibits 5´→3´ exonuclease activity
Direct to gel convenience
Flexibility to optimize Mg concentration in PCR
Supplied at a concentration of 5u/μl

The second generation of GoTaq products, GoTaq G2 Flexi DNA Polymerase reliably amplifies a wide range of PCR templates and provides high-performing results due to improved manufacturing processes, increased reliability and consistency. GoTaq G2 Flexi DNA Polymerase is supplied with 5X Green GoTaq Flexi Buffer and 5X Colorless GoTaq Flexi Buffer and 25mM MgCl2. The GoTaq G2 Flexi DNA Polymerases are supplied in a proprietary formulation containing 50% glycerol with buffers designed for enhanced amplification. The enzyme is a full-length form of Taq DNA polymerase that exhibits 5'-->3' exonuclease activity. The 5X Green GoTaq Reaction and Flexi Buffers contain two dyes (blue and yellow) that separate during electrophoresis to indicate migration progress. The colorless buffer is used when direct fluorescence or absorbance readings are required without prior purification of the amplified DNA from the PCR.

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GoScript Reverse Transcriptase Open/Close Add
Available as a standalone enzyme, a complete reverse transcription kit or as a master mix with Oligo(dT) or Random Primers
Sensitive transcription of both high-copy and low-copy messages
Transcribes short and long transcripts; processes through secondary structure

GoScript Reverse Transcriptase utilizes M-MLV and state-of-the-art buffer technology designed for qPCR to deliver robust, reliable cDNA synthesis of a full range of rare and abundant transcripts, even in the presence of inhibitors. GoScript Reverse Transcriptase is qualified for use in qPCR, including GoTaq qPCR and Plexor qPCR systems for performing RT-qPCR.

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dNTP Mix Open/Close Add
Highly pure (>99% triphosphate)
10mM each nucleotide in water
Reduce pipetting steps by adding nucleotides as a mix

dNTP Mix is a premixed solution containing sodium salts of dATP, dCTP, dGTP and dTTP, each at 10mM in water at pH 7.5; the total concentration of nucleotides is 40mM. One microliter of the dNTP Mix in a 50ul reaction will give a final dNTP concentration of 200uM for each dNTP.

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dNTP Mix Open/Close Add
Highly pure (>99% triphosphate)
10mM each nucleotide in water
Reduce pipetting steps by adding nucleotides as a mix

dNTP Mix is a premixed solution containing sodium salts of dATP, dCTP, dGTP and dTTP, each at 10mM in water at pH 7.5; the total concentration of nucleotides is 40mM. One microliter of the dNTP Mix in a 50ul reaction will give a final dNTP concentration of 200uM for each dNTP.

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Deoxyuridine Triphosphate (dUTP) Open/Close Add
Highly pure (≥99% triphosphate)
Supplied at 100mM in water for ease-of-use
Incorporate dUTP into PCR products and treat with uracil-N-glycosylase to inhibit subsequent amplifications

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5. dUTP (2'-Deoxyuridine, 5'-Triphosphate) can be used in place of dTTP in PCR and RT-PCR protocols to prevent carryover from previous amplifications. The substitution of dUTP for dTTP in PCR results in uracil-containing PCR products that are suitable for most standard applications. The enzyme uracil-N-glycosylase (UNG, also referred to as UDG) can be added to a PCR premix to excise uracil from any contaminating PCR product, thereby preventing false positives. Each lot of dUTP is function-tested to ensure specific DNA amplification and the absence of nuclease activity.

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Deoxynucleotide Triphosphates (dNTPs) Open/Close Add
Greater than 99% triphosphate content
100mM in water at pH 7.5

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5.

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Deoxynucleotide Triphosphates (dNTPs) Open/Close Add
Greater than 99% triphosphate content
100mM in water at pH 7.5

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5.

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AMV Reverse Transcriptase Open/Close Add
DNA polymerization using template DNA, RNA or RNA:DNA hybrids
Requires a primer as well as magnesium or manganese
Stable at higher reaction temperatures (37–58°C)
Supplied at a concentration of 10u/μl (Cat.# M5101, M5108) or 20–25u/µl (Cat.# M9004)

Avian Myeloblastosis Virus Reverse Transcriptase (AMV RT) catalyzes DNA polymerization using template DNA, RNA or RNA:DNA hybrids. It requires a primer (DNA primers are more efficient than RNA primers) as well as Mg(2+) or Mn(2+). The enzyme possesses an intrinsic RNase H activity. Both nonionic detergents and sulfhydryl compounds stabilize the enzyme activity in vitro.

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Access RT-PCR System Open/Close Add
All the reagents you need for single-tube reverse transcription and PCR
Separate components allow you to control every step of the reaction

The Access RT-PCR System is designed for the reverse transcription (RT) and PCR amplification of a specific target RNA from total RNA or mRNA. This one-tube, two-enzyme system provides sensitive, quick and reproducible analysis of even rare RNAs. The system uses AMV Reverse Transcriptase (AMV RT) from Avian Myeloblastosis Virus for first-strand DNA synthesis and thermostable Tfl DNA polymerase from Thermus flavus for second-strand cDNA synthesis and DNA amplification. The Access RT-PCR System includes an optimized single-buffer system that permits extremely sensitive detection of RNA transcripts without a requirement for buffer additions between the reverse transcription and PCR amplification steps. This simplifies the procedure and reduces the potential for contaminating the samples. In addition, the improved performance of AMV Reverse Transcriptase at elevated temperatures in the AMV/Tfl 5X Reaction Buffer minimizes problems encountered with secondary structures in RNA.

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