Question

Often in laboratory experiments you will want to amplify regions of the genome from several different preparations of gDNA. To do this, a mastermix will be created with all the common reagents to ensure easier pipetting, proper mixing, and controlling for variations in pipetting. Create a mastermix for running 8 identical reactions, including one negative control, each with different gDNA templates (7 total gDNA templates).

Answer 1

Mastermix in PCR helps in providing faster and easier setup with pipetting, proper mixing, reduction in contamination and controlling fir variation in pipeting or tube to tube variability. Contents of PCR master mix for the desired reaction are as follows:

• dNTPs
• Primers
• thermostable DNA polymerase
• cofactor (MgCl2)
• Template DNA (for 7 samples)

• PCR-grade water

The only difference in negative control for the PCR master mix is template or the different primer rest the components are the same in the master mix.