The normal (wildtype) size of
The normal (wildtype) size of the brat gene is 1500 basepairs (bp). In the mutant variant you identified, there is adeletion of 500 bps in the last exon of the gene. How would yourecognize the two versions based on size? Listwhat fragment sizes you would expect to see on a gel.
Answer:
The identification of the two variants can be done with PCRusing two different primer sets. As there is a deletion in the lastexon of the gene, primers designed for the amplification of thefull-length brat gene will not succeed in the amplification ofmutant variant. So another primer that binds to the sequence beforethe deleted fragment will be used. Below figure shows the strategyfor designing the experiment
Two PCR amplification will be done in each sample- wild typebrat and mutant brat using promer set P1/P2 and P1/P3. theresultant PCR amplification after gel run will be as below.
In the wildtype sample, there will beamplification in both the primer combination. However in themutant, because of the deletion of 500bp, the primer combinationP1/P3 will not lead to any amplification in the PCR and only thePCR combination P1/P2 will lead to amplification with amplicon sizeof 1000bp.
"Our Prices Start at $11.99. As Our First Client, Use Coupon Code GET15 to claim 15% Discount This Month!!"
