Focus: AUTOMATED NUCLEIC ACID AND PROTEIN PURIFICATION
Search the New Maxwell® 16 Applications Database for Specific Sample Information
Maxwell® 16 System offers automated walk-away nucleic acid and recombinant protein extractions and purification. The simple design of the Maxwell® 16 System and robust chemistry in the prefilled reagent cartridges allows processing of a wide variety of sample types. Here we introduce the Maxwell® 16 Applications Database, designed to allow users to share information about purification of nucleic acid or protein from specific sample types.
Promega Corporation
Published in October 2007
Introduction
The Maxwell® 16 Applications Database was developed to facilitate sharing of Maxwell® 16-related knowledge (www.promega.com/maxwell16/applications). The database contains information about samples tested by both Promega scientists and Maxwell® users throughout the worldwide scientific community. This database allows you to search by keyword or browse through folders (Figure 1) and contains information about samples processed, typical yields achieved and downstream applications tested. Join our community of Maxwell® users by submitting your data today at: www.promega.com/maxwell16/applications/submission.asp.
Figure 1. Maxwell® 16 Applications Database Advanced Search Window. The database can be accessed at: www.promega.com/maxwell16/applications.
The following are several recently submitted entries to the Maxwell® 16 Applications Database.
DNA from Formalin-Fixed, Paraffin-Embedded Mouse Tissues
DNA IQ™ Casework Sample Kit for Maxwell® 16 (Cat.# AS1210) was used to extract DNA from formalin-fixed paraffin embedded mouse intestine tissue sections (5 microns). The Tissue and Hair Extraction Kit (for use with DNA IQ; Cat.# DC6740) was used for preprocessing of samples. Sections were incubated with 1.8mg/ml Proteinase K and 100mM DTT in Incubation Buffer 16 hours at 56°C, then transferred to well #1 of the DNA IQ™ Casework Sample Cartridge. The LEV (Low Elution Volume) setting and Forensic method were used, and DNA was eluted in 100ul Elution Buffer. Eluted DNA was not detectable by spectrophotometry, but DNA was amplified by end-point PCR (Figure 2) and real-time PCR (data not shown).
Figure 2. Amplification of DNA Extracted Using the Maxwell® 16 Instrument from Formalin-Fixed, Paraffin-Embedded Mouse Tissue. DNA was extracted from 1, 3, and 5 five micron intestine sections after overnight Proteinase K digestion using the DNA IQ™ Casework Sample Kit for Maxwell® 16. Five microliters eluted DNA was amplified in 20ul PCR using GoTaq® Green Master Mix and beta actin primers. Amplification of DNA from sections of two different tissue blocks is shown.
DNA from Saliva
Maxwell® 16 Cell DNA Purification Kit was used to extract DNA from human saliva. Saliva (400ul) was transferred directly to well #1 of the Maxwell® 16 Cell DNA Purification Cartridge and processed following the SEV Cell DNA method. Saliva yielded 0.7–3µg DNA. Eluted DNA was amplifiable using human DNA-specific primers (Figure 3, Panel A) and eubacterial 16S ribosomal primers (Figure 3, Panel B).
Figure 3. Amplification of DNA Extracted from Saliva Using the Maxwell® 16 Instrument. Panel A: PCR with human Factor V primers (5µl template in 25µl total reaction, 30 PCR cycles). Panel B: PCR with Eubacteria 16S rRNA primers, (2µl template in 25µl total reaction, 30 PCR cycles). Lanes M, BenchTop PCR Markers (Cat.# G7531); lane 1, no template control; lane 2, human genomic DNA control (Cat.# G3041); lane 3; Sample A; Lane 4, Sample B; Lane 5, Sample C.
RNA from Adipose Tissues
Maxwell® 16 Total RNA Purification Kit was used to extract RNA from rat adipose tissue. Fifty to 150mg adipose tissue was processed per cartridge. Varying amounts of Clearing Agents were tested (0.5–1.5x), and half the standard recommended amount of Clearing Agent was found to give the best yield, likely due to the relatively low amount of RNA in the adipose tissue. RNA yields were ~2–4µg, and beta actin and GAPDH transcripts were detected by RT-PCR (Figure 4). No contaminating genomic DNA was detectable by real-time PCR (data not shown).
Figure 4. RT-PCR Analysis of Adipose Tissue Total RNA Extracted Using the Maxwell® 16 Instrument. RNA was analyzed by AccessQuick™ RT-PCR using beta actin primers (lanes 2-3) or GAPDH primers (lanes 5-6). Lanes 1 and 4, no-template control; lanes 2 and 5 input RNA purified from 100mg tissue; lanes 3 and 6 input RNA purified from 150mg tissue.
Summary
These and may more applications for the Maxwell® 16 Instrument are available at the click of your mouse from the online database. Search for information about a sample type that interests you, or submit your data for the benefit of others.