Below are the images of RNA gels Discuss what you see in eac

Below are the images of RNA gels. Discuss what you see in each lane (1, 2 and 3). Provide your assessment of RNA quality. Can each of these preps be used for quantitative RT-PCR? Explain: What types of problems can be expected using all these preps for qRT-PCR? Support your statements with your knowledge and prediction of what could affect your results in each case.

Solution

Lane 2 showing intact RNA with 28S (1st band) and 18S (2nd band) RNA in 2:1 proportion.

Lane 1 showing smear of degraded RNA, 28 S RNA is degraded and is accumulated in bottom.

Lane 3 showing slight contamination of genomic DNA (the first faint band near the well).

Degraded RNA can not be used for qRT PCR because it can not be reverse transcribed to cDNA by enzyme reverse transcriptase as this enzyme catalyzes cDNA synthesis starting with the polyA tail on mRNA.

Genomic DNA contamination can cause overestimation of the actual RNA concentration as it can be amplified along with cDNA during qRT PCR.

Lane 2 sample can be used .as it is intact RNA.