I have GeNei PCR Master Mix of 2x concentration shall I dilute it to 1x before using for PCR or use it directly without any dilution
Can someone tell me what is the best way to use Master Mix
Yes I agreed with Nahed Hussien
. Don't need to dilute the master mix. Your master mix is ready for use.Your total reaction volume sould be 25μl where 12.5μl goes master mix, 0.3-0.4μl primers, 1-2μl DNA sample (depending on yeld conc), and rest of the volume is fullfiled by nuclease free water.
Best wishes
2x PCR master mix means that it is 2 fold concentrated. As in the previous answers mentioned, you do not need to dilute the master mix prior use. You can directly set up your PCR reaction. You only need to keep in mind that half of the final volume of your reaction has to be your PCR master mix. Further you only need to add primers and template DNA and fill up with H2O to the desired final volume.
Best wishes
No, it does not need diluting. As explained above, when you set up the reaction, the proportion of master mix to other reagents makes the mix be 1X in the final concentration.
If you were to dilute the master mix to 1X prior to use, then the addition of primers and DNA would make the mix be less than 1X in concentration.
If you run PCR reactions infrequently or in small numbers I would suggest aliquoting the mix into smaller volumes before you freeze it for the first time. Repeated freeze/thaw cycles will reduce the activity over time.
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