those three steps and how do they facilitate the amplification of your products?
ID: 260284 • Letter: T
Question
those three steps and how do they facilitate the amplification of your products? b) Below is an example of a gel from a similar experiment. Analyze this gel and answer the questions. The lanes were loaded as follows: Lane I: DNA loading ladder Lane 2: Empty lane Lanes 3-5: Control Genomic DNA with actin primers Lane 6: Empty lane Lanes 7-9: Control Genomic DNA with hsp70 primers Lane 10: Empty lane Lanes 11-13: Control cDNA with actin primers Lane 14: Empty lane Lanes 15-17: Control cDNA with hsp70 primers 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 1. Why are the bands in lanes 3, 4 and 5 on the gel about 5) bps/larger than the bands in lanes 11, 12 and 13? Lanes 15, 16 and 17 on Gel are empty. Is this a mistake? Or is this expected? If it is , a mistake, offer an explanation. If it is expected, offer an explanation/ Draw a picture of your gay and your expected results. Make sure to label yaur lanes. IExplanation / Answer
Please find the answers below:
Answer 1: According to the image, the bands observed in the genomic DNA and the cDNA for actin primers have different locations. The bands with genomic DNA are slighlty higher in location with larger molecular weight as compared to the cDNA bands. This is because the whole genome segment in the lanes 3,4 and 5 have been amplified in this case. The amplification of the genomic DNA starts from a few base pairs higher than the usual and is extended upto a little longer than usual, thus increasing the size of the bands observed since the primers might bind a few base pairs above and below the target. Unlike the genomic DNA, the cDNA contains the exact sequence for amplification and hence gives a band of exact molecular weight.
Answer 2: The lanes 15-17 are empty but this is not an experimental mistake. The sample in these lanes utilizes cDNA and primers for hsp70 are used for amplification. The fact that the bands are not visible can be explained in two manners:
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