27845 Irma Lee Circle, Unit 101
Lake Forest, IL 60045
EzWay Direct PCR Buffer (5x)
EzWay™ Direct PCR Buffer enables the direct amplification of DNA fragments from a variety of samples including whole blood, blood collected on card, saliva, bacteria, mouse tail, tissue, cultured cells or plant eliminating the need for DNA purification, just by adding to the PCR mixture in place of general 10X PCR Buffer.
EzWay™ Direct PCR Buffer is compatible with most commercially available any DNA polymerases including Taq, Pfu, Pwo, Tth and even with any type of Hot Start Taq DNA polymerase. A one-step reaction using this EzWay™ Direct PCR Buffer enables easy PCR and minimizes the sample cross-contamination when dealing with multiple tissue or samples by eliminating tedious genomic DNA preparation.
6 months at 4°C, 1 year at -20°C
Fig.1 Genomic DNA PCR vs. Blood Direct PCR of different sizes of p53 targets
PCR amplifications were performed using 20 ng human genomic DNA or 1 ul Heparin-treated whole blood in a 50ul PCR reaction volume. EzWay™ Direct PCR Buffer can amplify directly up to 2kb amplicon and produce more specific band patterns than DNA PCR. It means that direct PCR amplification has inherent hot start function even without the help of hot start enzyme because the cells are disrupted in initial heating stage, then the exposed DNA templates meet primers instantly at higher temperature than Tm.
Fig.2 Direct PCR of p53 from human whole blood Blood treated with three anti-coagulants
Direct amplification from anticoagulants treated human blood with EzWay™ Direct PCR Buffer (Lane2-7) and conventional PCR buffer (Lane1). The inhibitory effects caused by anti-coagulants and blood components is successfully overcome and PCR result can be achieved even in the condition of maximum 10-20%(v/v) blood volume in a total reaction volume.
Lane P: Human genomic DNA, Lane N: Negative controls
Fig.3 Direct sequencing from DNA and EDTA-blood Apolipoprotein E genotyping
Apolipoprotein E, e2/e4, is identified with DNA (A) and EDTA-blood (B) by PCR and direct sequencing using BigDye® Terminator v3.1 cycle sequencing reaction. The amplicon from blood by EzWay™ Direct PCR Buffer can be used as sequencing template. Then, the direct PCR chemistry does not interfere downstream dideoxy sequencing reaction and fluorescent detection.
Fig.4 AmpliTaq Gold specific Direct PCR Buffer Gold
Blood having 5 different genotypes (Lane 1,6,11, Lane 2,7,12, Lane 3,8,13, Lane 4,9,14 and Lane 5,10,15, respectively) was amplified by Direct PCR method. The Direct PCR Buffer Gold is specific to AmpliTaq Gold DNA Polymerase as shown at the lane 6-10. AmpliTaq Gold didn't work well when it was used with general type of Direct PCR Buffer (lane 16-20).
Lane 1-5 : Direct PCR using Direct PCR Buffer Gold (K0568011) with KOMA Taq DNA Polymerase
Lane 6-10 : Direct PCR using Direct PCR Buffer Gold (K0568011) with AmpliTaq Gold DNA Polymerase
Lane 11-15 : Direct PCR using Direct PCR Buffer (K0568001) with KOMA Taq DNA Polymerase
Lane 16-20 : Direct PCR using Direct PCR Buffer (K0568001) with AmpliTaq Gold DNA Polymerase