Describe the ingredients necessary for a PCR reaction (such as the one in lab 8), and...

Describe the ingredients necessary for a PCR reaction (such as the one in lab 8), and why they are necessary.

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Expert Solution

Template DNA , DNA polymerase, primers, deoxy nucleoside triphosphate, buffer and Mg2+ cofactor and buffer are the 6 main components of PCR.

The template strands contain our target sequence which will be amplified . The genome can be from any source, a plasmid DNA, human genome , bacterial, viral etc. Always an optimum amount is better to increase specificity . Primers are oligonucleotide sequence which binds to flank regions of target DNA sequence. It is primer which decide the nature of target sequence. Primer is designed depending on our sequence weather to amplify virulant genes, drug resistant gene etc. They are 15-30 bp long

DNA polymerase is the enzyme which is required throughout.Taq polymerase is the enzyme used which withstand in high temprature. They provide thermaltability and add nucleotide at a temprature at 70°c.dNTPs are building blocks of new DNA strand . They are of 4 nucleotides added optimal base in operation. Excess incorporation may leads to inhibition or misincorperation of bases. Magnesium is essential cofactor for function of DNA polymerase which will enable dNTPs binding. It also help in formation of phosphodiester bond . It also facilitate complex between primers and template by stabilizing negative charge . Suitable chemical environment required of polymerase chain reaction is provided by buffer. Potassium ions of KCL helps in primer annealing. Sometimes pottasium is replaced by ammonium ions which will increase specificity by destabilizing effect. It helps in maintaining ionic imbalnce and helps in amplification.

Thank you.

gladiator answered 1 year ago

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