PCR MASTER MIX

ORDERING INFORMATION

	Catalog #	Number of 50 µl 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 225–300nm, 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 µl with water:

Component	Volume in µl	Final concentration
2x PCR-Master Mix	25	1x (1.5 mM MgCl2)
Forward-Primer	variable	0.2 – 1 µM
Reverse-Primer	variable	0.2 – 1 µM
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 dNTP’s 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

WARNING