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PCR MASTER MIX

ORDERING INFORMATION


Catalog #

Number of 50 l Reactions

Volume

Price (EUR)

2x PCR Master Mix

# R 500
100 2 x 1.25 ml 39,00
# R 501
500
10 x 1.25 ml 167,00
2x Red PCR Master Mix
# R 540
100
2 x 1.25 ml 42,75
# R 541
500
10 x 1.25 ml 170,00
2x Green PCR Master Mix
# R 570
100
2 x 1.25 ml 42,75
# R 571
500
10 x 1.25 ml 170,00
2x Hot Start PCR Master Mix
# R 530 100
2 x 1.25 ml 56,00
# R 531 500
10 x 1.25 ml 234,00
2x Red Hot Start PCR Master Mix
# R 520 100
2 x 1.25 ml 74,00
# R 521 500
10 x 1.25 ml 305,00
2x Green Hot Start PCR Master Mix
# R 580 100
2 x 1.25 ml 74,00
# R 581 500
10 x 1.25 ml 305,00


DESCRIPTION

The 2x PCR Master Mix contains all reagents required for PCR and is designed to make PCR as easy and simple as possible. All components (inclusive Taq DNA Polymerase respectively Hot Start DNA Polymerase) are provided in an optimized concentration in the 2x PCR-Master solution. With 2x PCR Master Mix all you need to do is to add primers and template DNA, thus minimizing the pipetting effort and possible sources of error. The mix is suitable for PCR amplification of DNA-fragments up to 4 kb, in most cases even longer targets can be successfully amplified.

Red PCR Master Mix and Red Hot Start PCR Master Mix contain an inert red dye which allows identification of the reactions which contain enzyme. The dye has no adverse effect on PCR.

Green PCR Master Mix and Green Hot Start PCR Master Mix contain two dyes (blue and yellow) that separate during electrophoresis to monitor migration progress. Reactions assembled with Green PCR Master Mix have sufficient density for direct loading onto agarose gels. Green Master Mix is recommended for any amplification reaction that will be visualized by agarose gel electrophoresis and ethidium bromide staining. The dyes absorb between 225300nm, making standard A260 readings to determine DNA concentration unreliable.


QUALITY CONTROL ASSAYS

FUNCTIONAL ASSAYS

SPECIFICATION

4 kb PCR

suitable

PHYSICAL ASSAYS

SPECIFICATION

Endonuclease Assay

no activity detected

DNase Assay

no activity detected

RNase Assay

no activity detected


BASIC PROTOCOL

Combine the following components in a PCR-reaction tube and adjust to a final volume of  50 l with water:

Component

Volume in l

Final concentration

2x PCR-Master Mix

25

1x (1.5 mM MgCl2)

Forward-Primer

variable

0.2 1 M

Reverse-Primer

variable

0.2 1 M

Template DNA

variable

1 150 ng

Distilled, sterile water

Add to a final volume of 50


Final volume

50


Mix gently and place in thermal cycler. No vortexing, no centrifugation. Optimal conditions for concentration of primer, template and temperature profile need to be determined for each reaction.


TROUBLESHOOTING

Observation

   Check



Nonspecific products (smearing)

  • Concentration of enzyme, primer, and/or dNTPs too high
  • Annealing temperature for primers too low
  • Too many cycles
  • Annealing and extension time too long
  • Too much template DNA


Low yield of product

  • Not enough or to much enzyme
  • Denaturation/extension temperature too high
  • Incorrect annealing temperature
  • Too few cycles
  • Poorly designed primers
  • Inhibitors from DNA purification (i.e. SDS)


No product

  • Incorrect annealing temperature
  • Incomplete denaturation
  • Poorly designed primers
  • Use of destroyed components due to wrong storage

STORAGE

Store at -20C for 12 months. Multiple freeze-thaw-cycles should be avoided by preparing aliquots.


WARNING

For research only. Not recommended or intended for diagnosis of disease in humans or animals. Do not use internally or externally in humans or animals.