Sequence 1: 5'ACTGTCGATGCTAGCTTGATCCAAGTATTGCTAGACAGAATTGACATATAGGCGATGCTAGT3' Sequence 2: 5'ATCGCTAGGATCGCTAGATTTAAGTCGCTGATCGGCTAGATATAACAGGTCCTGAATCGCTA3' Design a splint oligomer and write the all-possible content(s) needed...

Sequence 1: 5'ACTGTCGATGCTAGCTTGATCCAAGTATTGCTAGACAGAATTGACATATAGGCGATGCTAGT3'

Sequence 2: 5'ATCGCTAGGATCGCTAGATTTAAGTCGCTGATCGGCTAGATATAACAGGTCCTGAATCGCTA3'

Design a splint oligomer and write the all-possible content(s) needed for reaction to occur.

Solutions

Expert Solution

Splint Oligomers: These are the nucleotide sequences that are use to join two consecutive sequences. This is an in-vitro approach where two double stranded break DNA strands are joined using these oligomers.

The rquired components are:

A DNA double stranded break strand
A synthetic Splint oligomer
Polynucleotide kinase
Radio labeled P32ATP
T4 DNA ligase

Here the minimum sequence of splint oligomer that is required is tetramer but it should have few G:::C base pairing as it is important for splint oligomer to hold the two sequence tightly together. As G:::C provide strong bonding.

Splint sequence:

Sequence 1: Sequence 2: 5'ACTGTCGATGCTAGCTTGATCCAAGTATTGCTAGACAGAATTGACATATAGGCGATGCTAGT3' 5'ATCGCTAGGATCGCTAGATTTAAGTCGCTGATCGGCTAGATATAACAGGTCCTGAATCGCTA3'

3' CGATCAATCTAGCG5' Splinter oligomer

First the Polynucleotide kinase will add the Radio labeled P32ATP at 5' end of 5'ATCGCTAGG (Sequence 2).

This will proceed by the phosphate diester bond of attack of radio labeled P32ATP at 5' end of 5'ATCGCTAGG by the nucleophilic 3'-OH end of sequence 1.

The DNA ligase fill facilitate the bond formation.

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gladiator answered 1 year ago

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