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EzWay Direct Taq PCR MasterMix (2x)
EzWay Direct Taq PCR MasterMix (2x)
Catalog NumberK0568010
PriceLogin
Size1ml
TypePCR Kit
Manual/Data Sheet
* This manual on-line is just for reference. The contents of this datasheet are subject to change from lot to lot.
Description

EzWay™ Direct Taq PCR MasterMix is a premixed solution containing Taq DNA Polymerase, dNTPs, MgCl2 and PCR Direct Buffer in a 1.5ml tube.
EzWay™ Direct Taq PCR MasterMix provides robust PCR performance in a wide range of PCR applications without the need for time-consuming optimization. Only primers and template DNA are added to prepare the final PCR.

EzWay™ Direct Taq PCR MasterMix allows direct PCR from all types of anticoagulant-treated bloods (heparin, EDTA, sodium citrate), plasma, serum, buffy coat, buccal swab without any DNA purification steps.
Also, EzWay™ Direct Taq PCR MasterMix can be used for direct PCR of various samples such as tissues, paraffin-embedded tissue slide or array, cultured cells, body fluids, bacteria, viruses, plants, yeast, hairs and so on.
According to experimental design, sample itself or crude lysate or DNA can be can be used as template for direct PCR. Especially, sample itself as template give naturally occurring "hot start PCR" effect without manual hot start method or hot start PCR enzyme.

Features
  • No need to purify DNA prior to PCR
  • Minimizes the risk of DNA contamination during reagent handling
  • Prevents the loss of trace samples in the DNA purification process
  • Amplification of DNA directly from samples containing : Whole Blood, Blood Stains, Blood Cards, Buccal Swab, Saliva, Hair root, Sperm, Body Fluid, Cultured Cells, Bacteria, Viruses, and Tissues (fresh, frozen or paraffinized), Plant etc.
  • Compatible with various sources of thermostable DNA polymerases
Applications
  • Viral infection / Molecular diagnostic test
  • Forensic DNA Analysis
  • Single cell diagnostic
  • Blood banking
  • Identity testing
  • Multiplex PCR / SNP detection / PCR-RFLP
  • Sequencing / Cloning
  • Laboratory Automatic PCR
Storage
6 months at 4°C, 1 year at -20°C
Packing
Component Cat.No Volume Comment
2X EzWay™ Direct Taq PCR Master Mix* K0568010 or K0568020 1.0 ml Final 1.5mM MgCl2 0.2mM dNTP 1U Taq / 20ul PCR with or without red-dye
4X Magic Buffer K0561031 1.0 ml For amplification of high GC contents
Note:
For the application of real time PCR, K0568020 must be used.
Figure 1. Direct PCR amplification of p53 exon6 from human fresh blood treated with anticoagulants
Direct PCR amplification of different sizes of p53 targets from heparin-treated human blood
PCR was performed using 20ug of both human genomic DNA and 1ul heparin-treated whole blood using EzWay™ Direct PCR MasterMix. It can amplify directly up to 2kb amplicon and produce more specific band patterns than PCR of DNA smaple. It shows that direct PCR amplification has inherent hot start function because the cells are disrupted in the initial heating stage, then the exposed DNA templates meet primers instantly at higher temperature than Tm.
Figure 2. Direct PCR amplification of different sizes of p53 targets from heparin-treated human blood
Direct PCR amplification of different sizes of p53 targets from heparin-treated human blood
PCR was performed using 20ug of both human genomic DNA and 1ul heparin-treated whole blood using EzWay™ Direct PCR MasterMix. It can amplify directly up to 2kb amplicon and produce more specific band patterns than PCR of DNA smaple. It shows that direct PCR amplification has inherent hot start function because the cells are disrupted in the initial heating stage, then the exposed DNA templates meet primers instantly at higher temperature than Tm.
Figure 3. Bovine melanocortin receptor 1 (MC1R) gene polymorphism test from beef meat by direct PCR-RFLP
Bovine melanocortin receptor 1 (MC1R) gene polymorphism test from beef meat by direct PCR-RFLP
50ng DNA purified from beef meat, 1ul of 1mg beef meat crude lysate in 100ul lysis solution, and 1ul of 10mg beef crude lysate in 200ul lysis solution were directly amplified without any purification step(lane 1,3,5) and totally cut by restriction enzyme(lane 2,4,6).
Figure 4. EGFR (Epidermal Growth Factor Receptor) mutation detection (exon 18-21) from paraffin-embedded tissue section of patients with NSCLC by direct PCR
EGFR (Epidermal Growth Factor Receptor) mutation detection (exon 18-21) from paraffin-embedded tissue section of patients with NSCLC by direct PCR
After tissue was deparaffinized, tumor region was scratched and the power was added directly to PCR mixture. All PCR reaction was performed with 25 ul PCR volume using Ezway™ Direct PCR MasterMix.
EGFR Exon 12 (Lane 1)/Exon 19 (Lane 2)/Exon 20 (Lane 3)/Exon 21 (Lane 4) amplified products were extracted from gel and sequenced and compared with matched normal blood to know the somatic mutation of tumors.
Citation List
  • Optimizing a qPCR Gene Expression Quantification Assay for S. epidermidis Biofilms: A Comparison between Commercial Kits and a Customized Protocol A França, AI Freitas, AF Henriques PLoS ONE, 2012 (abstract)
  • Efficacy of peginterferon and ribavirin is associated with the IL28B gene in Korean patients with chronic hepatitis C SH Jeong, YK Jung, JW Yang, SJ Park, JW Kim Clinical and Molecular Hepatology, 2012 (abstract)
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