What volume should be used of 2X Mastermix ?

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Manoj Pohare

For 25 microL of reaction you need to use 12.5 microL of 2X PCR master mix and yes you need to adjust the final concentration of PCR master mix to 1X.

Duaa - Duaa

thankx Manoj.can you guide me further for how to adjust it ti 1x?

Its simple. Prepare your reaction mixture (template, forward and reverse primers) and then add water so that final volume of the reaction become 12.5 microL. To it add 12.5 microL of 2X PCR master mix. In this way your 25 microL PCR reaction is ready with final concentration of PCR master mix to be 1X.

Best luck.