Problem3 - use conditions that will allow hybridization of...

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MCB 161 S. Harmer Problems for Lecture 3 – Molecular Methods MBOC Problems book: 4-55, 8-67, 8-80 ( note there is a typo in the book; you should be able to derive the correct answer nonetheless ), 8-87, 8-95 1. You wish to perform a Southern blot using as a probe a 1200 bp fragment of an actin gene (50% G + C). You know that the genomic DNA sample that you are going to hybridize to includes several actin genes. One is 100% identical in sequence to your probe. The others are at most 80% identical (20% mismatch) to your probe. You want to
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Unformatted text preview: use conditions that will allow hybridization of your probe to the identical gene, but will prevent hybridization of your probe to the other genes. The only incubator you have is stuck at 55C so you will have to perform the hybridization at this temperature. Determine a set of conditions (salt concentration (Ci) and % formamide) that will be appropriate for this hybridization. 2. See the link to the BLAST question on the Lecture Info course web page....
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