Questions 13-15 apply to the scenario below A ßB2-crystallin gene knockout mouse

Question

Questions 13-15 apply to the scenario below A ßB2-crystallin gene knockout mouse was produced by replacing part of the Crybb2 gene with a neomycin resistance gene (NeoR). To confirm the knockout, DNA was extracted from knockout and wild-type mice, and PCR amplification with appropriate primers (primers for Crybb2 and NeoR) was performed. 13- Based on the results obtained after running your amplified samples on an agarose gel, which samples were extracted from the knockout mice? A. 1 and 4 B. 3 and 4 1000bp- 750 te 500 bp

Explanation / Answer

Genotyping is performed using two sets of primers.
1. Gene-specific primers
2. Insertion (Mutation) specific primers
WT organism gives a PCR product only with the gene-specific primers and not with the mutation-specific primers.
Mutant organism gives a PCR product only with the mutant-specific primers and not with the WT specific primers.
A heterozygous individual produces both the bands specific to WT PCR product and mutant PCR product.

In the given case,
The knockout mice should produce a band specific to the NeoR gene and it should not produce a band specific to the crybb2 gene.
Lane 1 displays a band specific to the NeoR gene and Lane 2 does not display a band specific to the WT crybb2 gene.
So, both these lanes belong to the homozygous knock out mouse.

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