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TP SYBR 2x Master Mix

Description

TP SYBR 2x Master Mix is dedicated for universal analysis of DNA samples using qPCR with quantification of DNA amplicons with DNA dye SYBR Green I. It is based on recent finding that addition of Trehalose or 1,2-Propanediol (abbriated TP) into reaction mixture is capable of susbstantialy increasing efficiency of PCR and enhance amplification of samples which are otherwise difficult to amplify, including DNA from whole blood, GC rich amplicons and samples containing PCR inhibitors (Horáková a spol., BMC Biotechnology, 11:41, 2011; Free PMC article).

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SYBR green I

  • The Mix contains fluorescent DNA dye SYBR Green I, which after binding to double stranded (ds)DNA becomes strongly fluorescent with maximal excitation at 497 nm (blue light) and emission at 520 nm (green light). Because fluorescence of unbound SYBR Green I is low, enhanced fluorescence during qPCR corresponds to an increase in dsDNA amplicons produced during PCR.

Rapid preparation of the samples

  • All components of the TP SYBR 2x Master Mix are 2x concentrated (optimized reaction buffer containing trehalose and 1,2-propanediol, nucleotides, Taq DNA polymerase, SYBR Green I) which facilitates rapid preparation of the samples. The samples are prepared by mixing an aliquot of the Master Mix with oligonucleotide primers, template DNA and H2O (included).
  • TP SYBR 2x Master Mix is especially useful for routine analyses of large numbers of DNA samples. To 0.5 ml of the Master Mix in original tube, primers (e.g. 40 µl forward and 40 µl reverse) and PCR H2O (e.g. 380 µl) are added and mixed; the "armed" Mix can be stored at -20 ± 5oC. Immediately before use, the Mix is thawed and each e.g. 24 µl aliquot is mixed with e.g. 1 µl of the tested DNA template.

 

Technical data

Components and basic packaging

  • 1 tube with 0.5 ml TP SYBR 2x Master Mix (for 40 reactions, 25 µl each). 
  • 1 tube with 1.5 ml PCR Ultra H2O (in basic package).

Composition

  • TB SYBR 2x Master Mix contains: 150 mM Tris-HCl, pH 8.8 (at 25°C), 40 mM (NH4)2SO4, 0.4 M trehalose, 2 M 1,2-propanediol, 0.02% Tween 20, 5 mM MgCl2, 400 µM dATP, 400 µM dCTP, 400 µM dGTP, 400 µM dTTP, 100 U/ml Taq DNA polymerase, SYBR Green I, stabilizers and additives.

Storage

  • At temperature -20°C ± 5°C. Material can be repeatedly defrosted. For short period of times (up to 3 days) material can be stored at up to 30°C . This allows its sending at ambient temperature (nature friendly).

Purity and quality control

  • Purity of Taq DNA polymerase is verified by SDS PAGE, only one band of 94 kDa is observed in Coomassie blue stained gel. Material is free of nucleases.
  • Each batch of the TP SYBR 2x Master Mix is tested for amplification of a single copy gene in genomic DNA. 
 

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Cat. No. Product name and specification Amount Price
T605 TP SYBR 2x Master Mix 40 reactions 19.58 EUR
T606 TP SYBR 2x Master Mix 200 reactions 78.33 EUR
T607 TP SYBR 2x Master Mix 1000 reactions 313.33 EUR
T607xl TP SYBR 2x Master Mix 4x 1000 reactions 787.08 EUR