To further analyze the CRABS CLAW gene (see Problems 19 ...

Question

To further analyze the CRABS CLAW gene (see Problems 19 and 20), you create a map of the genomic clone. The 11-kb EcoRI fragment is ligated into the EcoRI site of the MCS of the vector shown in Problem 18. You digest the double-stranded form of the genome with several restriction enzymes and obtain the following results. Draw, as far as possible, a map of the genomic clone of CRABS CLAW.

EcoRI 11.0, 3.0 XbaI 4.5, 9.5
EcoRI + XbaI 4.5, 6.5, 3.0 XhoI 13.2, 0.8
EcoRI + XhoI 10.2, 3.0, 0.8 SalI 6.0, 8.0
EcoRI + SalI 6.0, 5.0, 3.0 HindIII 12.0, 1.5, 0.5
EcoRI + HindIII 9.0, 3.0, 1.5, 0.5

What restriction digest would help resolve any ambiguity in the map?

Accepted Answer

Welcome back everyone. Let's look at our next question. It says which of the following is a laboratory technique that involves using restriction enzymes to cut DNA at specific locations. Choice a restriction digest B PC R polymerase chain reaction C gel, electrophoresis or D DNA hybridization. Well, we've got a pretty big clue here in that. Our question tells us the technique we're looking for involves using restriction enzymes to cut DNA. And we have choice a restriction digest. And indeed, that is our answer. A restriction digest is when you use restriction enzymes to cut DNA. And of course, those restriction enzymes have specific cutting sites. They can leave blunt or sticky ends and can be used for all different sorts of things that you might want to cut your DNA leaving very specific ends. So it can be used for cloning, running things on gels and looking at uh different fragments, all sorts of activities. We'll just take a quick look through our other answer. Choices. Choice B PC R is used to amplify DNA segments. So you have a specific DNA segment, you'd like many copies of, you use primers on either end, run it through PC R and you can go from a very tiny little sample to having large amounts of your desired DNA sequence. But that's not what we're looking for. So, Choice B is not correct to gel, electrophoresis is of course a method of separating molecules by size, running an electric current through a gel and then the molecules migrate through that through the pores of the gel, but not what we're looking for. So choice C is not our answer and finally choice D DNA hybridization causing a DNA strand to anneal to its complementary strand or to a complementary R N A strand. And often this is used uh in the use of complementary probes, you can label a probe with radioactivity or a color and let it find its complementary sequence and use that to find specific DNA sequences but not what we're looking for here. So choice D is not correct either. So again, the laboratory technique that uses restriction enzymes to cut DNA choice a a restriction digest. See you in the next video.

Key Concepts

Restriction Enzymes

Genomic Mapping

Ambiguity Resolution in Mapping

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