2xPCR mastermix - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki

Using a PCR master-mix saves time and improves PCR reproducibility, especially if many reactions are prepared. For a standard PCR reaction, half of the final volume should be used, for example 25 µL 2xPCR mastermix for a 50 µL PCR reaction. Scale up or down as needed.

Component	µL	stock	2xPCR mastermix	final concentration in PCR reaction
Water	16	-	-	-
Buffer(xC)	5	10x	2x	1x
MgCl2(mM)	2	50 mM	4 mM	2 mM
dNTPs(mM)	1	10	0.4	0.2
Taq	1	100%	5.0%	2.5%
Total	25	-	-	-

See standard pcr protocol.

The 2xPCR mastermix can be combined with a PCR compatible loading buffer such as the in described in the table below. This saves time if you have many samples as they can be loaded directly on a gel. Use 33 µL of the 1.5xGreen PCR mastermix in a 50 µL PCR reaction. Scale up or down as needed.

Component	µL
2xPCR mastermix	25
[[6 x DNA loading buffer (PCR compatible)	DNA-loading-buffer]]
Total	33

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