Describe the ingredients necessary for a PCR reaction, and why they are necessary.

Expert Solution

Polymerase chain reaction (PCR) :-
• This is in vivo amplification of DNA sample.
In this process, a DNA sample amplified with help of various ingredients which are -

1. Target DNA/ DNA sample -
• This is the DNA sequence which will undergo amplification process.
• Sample DNA strands are used as template strands in PCR. (* Both strands of sample DNA acts as template strands).

2. Primer :-
• Two primers or one set of primer is used in PCR.
• These are oligonucleotide sequences which determine from where the replication process will start.
• Primer provides attachment site for DNA polymerase enzyme (taq polymerase enzyme).

3. deoxynucleotide triphosphates (dNTPs) :-
• These are building blocks of DNA and used in formation of new strand during PCR.

4. DNA polymerase :-
• In PCR, a thermostable DNA polymerase is used, because this enzyme can survive on high temperatures (94-96°C) during PCR.
• This enzyme adds new nucleotides and forms new strand of DNA on template DNA.
• A specific thermostable DNA polymerase named taq polymerase is used in PCR.

5. Magnisium ions -
• These are cofactor for enzyme DNA polymerase.

6. Buffer solution -
• This solution maintains the pH of the reaction solution and makes pH ideal for enzymatic activity.

gladiator answered 2 years ago

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