Question

In: Biology

Comment on the suitability of the following pair of primers in amplifying target sequence of chromosomal...

Comment on the suitability of the following pair of primers in amplifying target sequence of chromosomal DNA.

(i) Forward primer: 5’ GCCTCCGGAGACCCATTGG 3’ Reverse primer: 5’ TTCTAAGAAACTGTTAAGG 3’

(ii) Forward primer: 5’ TCGAATTGCCAATGAAGGTCCC 3’ Reverse primer: 5’ GGGACCTTCATTGGCAATTCGA 3’

Solutions

Expert Solution

The properties of a good primer includes:
1. GC content between 40 to 60% the base pair ending must be with G or C
2. A melting temperature should be between 65 °C to 75 °C
3. The length of base pairs between 10 to 30 base pairs
4. For effective results, add 3 to 4 base pairs at 5' end at primer for restriction enzyme

i) Forward primer: 5’ GCCTCCGGAGACCCATTGG 3’ Reverse primer: 5’ TTCTAAGAAACTGTTAAGG 3’
In the forward primer it has a high G/C ratio and it at 3' end G G is present so this is an effective primer.
In the reverse primer, it does not contains significant amounts of G/ C ratio so this reverse primers might not work efficiently
(ii) Forward primer: 5’ TCGAATTGCCAATGAAGGTCCC 3’ Reverse primer: 5’ GGGACCTTCATTGGCAATTCGA 3
In the forward primer it has a high G/C ratio and it at 3' end G G is present so this is an effective primer.
In the reverse primer, it contains significant amounts of G/ C ratio so this reverse primers work efficiently


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