ALLin™ Hot Start Taq Mastermix, 2X

Applications


  • Hot-start PCR up to 6 kb, low copy target detection

  • PCR of complex (GC rich) templates

  • Fast PCR, multiplexing, TA cloning

Benefits

Catalog. # Product Name Size Price Qty
HSM0201 ALLin™ Hot Start Taq Mastermix, 2X 200 r of 50 µl
€189.50
HSM0205 ALLin™ Hot Start Taq Mastermix, 2X 1000 r of 50 µl
€852.50

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Description

highQu ALLin™ Hot Start Taq DNA Polymerase is the superior sensitive enzyme The activity at room temperature is blocked by small molecular inhibitor. Enzyme becomes active only after heating what allows for highly specific and extremely sensitive amplification, no primer dimer formation and no background.

In combination with the optimized ALLin™ buffer enzyme provides higher success rates in demanding PCR applications like amplification of complex or longer templates and fast cycling.

ALLin™ Hot Start Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, 4.5 x 104 (nucleotides incorporated before the error occurs) and produces A-tailed products suitable for ligating into TA cloning vectors.

The convenience of ALLin™ Hot Start Taq DNA Polymerase is maximized by the use of 2X Mastermix providing the additional advantage of reduced pipetting and minimized errors. The mastermix is even supplied with PCR Water, and the only thing to add is the template with primers.

Protocols

Print version of the protocol: Product Insert ALLin™ Hot Start Taq Mastermix, 2X
Important Notes
  • Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips.
  • Include a no-template control and positive control in parallel.
  • Thaw and keep reagents on ice. Mix well before use.
  • The longer the amplicon, the longer the extension time: 15 sec/kb - <6kb and 1 sec/kb - <1kb. Use >90 sec extension for multiplexing.
  • Run an annealing temperature gradient from 55°C to 65°C to choose the best specificity conditions.
  • Do not use fast cycling for multiplexing.
 
Prepare a 50 µl PCR reaction

 

Rev. & For. Primers variable, up to 0.4 µM final each (2 µl of 10 µM each)
cDNA Template or
gDNA Template
<100 ng or
5 - 500 ng
PCR Water to 25 μl
ALLin™ Hot Start Taq Mastermix, 2X 25 µl

 

  • Mix gently, avoid bubbles.
  • Place into the instrument set like:

 

Initial denaturation 1 cycle: 95°C – 1-2 min
Denaturation 40 cycles: 95°C - 15 sec
Annealing 40 cycles: 55-65°C – 15 sec
Extension 40 cycles: 72°C – 1- 90 sec (15 sec/kb)

 

  • After the PCR store probes on ice shortly, for long term storage, keep at -20°C.

Specifications

CAT. SIZE COMPONENTS COMPOSITION
HSM0201 200 r
of 50 µl
5 x 1 ml - ALLin™ Hot Start Taq Mastermix, 2X
5 x 1 ml - PCR Water
1X mastermix contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers.
HSM0205 1000 r
of 50 µl
25 x 1 ml - ALLin™ Hot Start Taq Mastermix, 2X
25 x 1 ml - PCR Water
1X mastermix contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers.


Storage


In the dark at -20°C.

Resources

Product Insert