HELP ASAP: Northern Blot ( the sizes of the bands and a description of the blot

Question

HELP ASAP: Northern Blot ( the sizes of the bands and a description of the blot is given) . zoom in screen to see blot. Please just read description below.

Northern Blot with ssRNA marker and random, non-specific probe binding. Lane 2 (biotinylated ssRNA ladder). Lane 4 (extracted mRNA)-liver tissue, lane 6 (tRNA liver tissue, lane 8 (tRNA)spleen tissue.

thick bands at 7000nt, 5000nt, and 3000nt.

Half of gel it there is random probe binding (seen as darker areas) in a diagonal line.

What is the significance of these results, Please explain in detail to understand the significance the results in the northern blot. If not going to be specific, then please allow someone else to answer. 1-3 sentences is not enough

Explanation / Answer

According to the information and image, it can be clearly understod that hepatic mRNA and tRNA are subjected to Northern blot analysis for identification of specific set of sequences upon them. Northern blot is a technique which utilizes complementary binding of the probe RNA to the target mRNA or tRNA and detects for presence of positive bands.

Here, it must be carefully noted that the lanes containing the mRNA and tRNA failed to show any significant bands. This clearly suggests that the random primer did not bind to either mRNA or tRNA extract from the liver and hence suggests two main parameters:

Thus, these are some of the reasons why a specific set of bands was not observed in any case. However, non-specific background was seen in the gel which demonstrates that the binding conditions of the probe need to be optimized for obtaining high resolution in the results.

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