8. (7 points) Short answers to questions: a. Compare and contrast Northern Blot

Question

8. (7 points) Short answers to questions: a. Compare and contrast Northern Blot and qReal-Time RT PCR techniques in following aspects: Which technique is more sensitive? Which technique displays a direct detection of RNA from the sample? Which technique may produce false positive signal? Why? When using qReal-Time RT PCR, how do we determine exact concentration of our test RNA? b. Reporter genes help us to reveal expression levels and patterns at the protein level. Several GFP transgenic mutant mice were designed: mouse #1 is expressing GFP in the eye tissue mouse #2 is expressing GFP in the skin tissue mouse #3 is expressing GFP in the heart and skin tissue mouse #4 was expressing GFP during embryonic stages only and not expressing GFP post-birth. Describe the general design/composition of the GFP reporter transgenic constructs for each mutant mouse

Explanation / Answer

Answer 8 a i) qRT-PCR is more sensitive as it can detect even the very less material via its amplification step, while northern blot becomes unable to detect such small quantities.

ii) Northern blot displays a direct detection of RNA from the sample.

iii) It's possible to get False signals in case of qRT-PCR when our primers become unable to be stretch over the splice sites/exon junctions in the genomic DNA.

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