Question

polymerase chain reaction to amplify a 500 bp region of human dna corresponding to a gene of interest. when your pcr reaction is complete, you plan to run the product on an agarose gel to determine if your pcr reaction worked correctly.

1. Explain how you going to make the pcr soup and why you going to make it this way.

2. State the what's in the control and why the controls are so important

Answer 1

1. PCR = Polymerase chain reaction
It is an in vitro DNA amplification method.

Steps:
i. Denaturation
ii. Annealing
iii. Extension
iv. Repeat

Components:
i. Template DNA
ii. Primers
iii. Buffer
iv. Taq polymerase
v. Water
vi. dNTPs

The PCR master mix is prepared in the following manner
i. For a 20 uL reaction, take 15 uL of deionized water
ii. Add 2 uL of 10X buffer
iii. Add 0.5 uL of 10 mM dNTPs
iv. Add 0.5 uL of Taq polymerase (1 unit)
v. Add 2 uL of primers ( 1 uL each forward primer and reverse primer)

2. A positive control contains a template DNA on which primers are working well. The absence of a band in positive control shows that there is a problem with reaction components.
This ensures that all the components of the reaction are in working condition.

A negative control contains all the components except primers.
This ensures that none of the components are contaminated. A band in negative control shows that reaction components are contaminated.

Please provide a POSITIVE RATING. Thank you