NXT Taq PCR Kit

Catalog No. 1108

Quick Overview

Hot Start PCR Kit designed for fast-cycling PCR on any thermal cycler, without affecting yield or performance

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$50.00

Details

Reagents Supplied

NXT Taq PCR Master Mix (2x) containing:

  • Hot Start NXT Taq DNA Polymerase 
  • Reaction Buffer
  • MgCl2
  • dNTPs

Water, Nuclease-free
10X Color Load Solution

Description

  • Allows to obtain a wide range of PCR products up to 4 kb and 10 kb from episomal or complex genomic DNA, respectively
  • Ready-to-use solution containing hot start NXT Taq DNA Polymerase, Reaction Buffer, MgCl2 and dNTPs
  • Designed for fast-cycling PCR on any thermal cycler
  • Shortens PCR cycling time without affecting the yield or PCR performance
  • Does not require redesigning of primers
  • Supplied with 10X Color Load solution which allows for direct loading PCR reactions on the gel
  • Replicates DNA at 72°C and exhibits a half-life of 40 minutes at 95°C
  • Contains the 5'→3' exonuclease activity
  • Lacks the 3'→5' exonuclease activity
  • Adds extra "A" at the 3' ends
  • Anti-Taq antibodies inhibit polymerase activity at moderate temperature
  • The polymerase activity is restored during the initial denaturation step.

Applications

  • Formation of complexes between Taq DNA Polymerase and an anti-Taq antibody forms a basis for Hot Start PCR, which allows for convenient room-temperature reaction setup.
  • Hot Start PCR may increase specificity, sensitivity and yield of a PCR reaction in comparison to the conventional PCR assembly method.

Notes

  • Do not use reaction volumes larger than 20 μl as this will interfere with the optimal temperature gradient required for successful results
  • Thaw, gently vortex and briefly centrifuge NXT Taq PCR Master Mix (2x), primers, DNA template before use to avoid localized differences in salt concentration
  • The annealing and extension steps require 5 s and 3 s per 100 bp, respectively
  • Set up PCR reactions at room temperature
  • Primers can be added separately or as a primer mix prepared previously
  • Vortex the samples and briefly spin down
  • Reactions can be placed in thermal cycler at room temperature
  • Use of 10X Color Load Solution allows PCR reactions to be loaded directly on the gel without prior addition of a gel loading buffer. 10X Color Load Solution contains a gel loading reagent and two tracking dyes (a red dye and an yellow dye) that separate during electrophoresis. The red dye migrates at the same rate as 600 bp DNA fragment and the yellow dye migrates faster than 20 bp in a 1% agarose gel. The dyes do not interfere with most downstream enzymatic applications, however it is recommended to purify PCR products prior enzymatic manipulation.
  • In most cases there is no need to add additives to the PCR reaction. For some difficult targets such as: GC-rich sequences, sequences with complex secondary structures additives such as DMSO can be added to improve amplification. Use DMSO in concentrations of 2-8%. The recommended starting DMSO concentration, if needed, is 3%
  • As a general guide for how much template DNA to use, start with a minimum 104 copies of the target sequence to obtain a signal in 25-35 cycles (i.e. 1 μg of 1 kb ds DNA equals 9.1 x 1011 molecules, 1 μg of E. coli genomic DNA equals 2 x 108 molecules, 1 μg of human genomic DNA equals 3 x 105 molecules)

Storage Conditions

Store @ -20°C
Product shipped on dry ice

Categories

Polymerases, Thermophilic DNA Polymerases