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recognition sites and cloning?

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#1 boa



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Posted 01 October 2012 - 08:12 PM

Hello all, I have absolutely no clue how to start this problem and I would like to have some cues or just something to chew on...thank you!!
I know what a plasmid is , I know what sticky ends (cohesive extremities) are but I dont know how the multiple cloning sites in this plasmid are located so im a bit lost!

Question: U must clone a DNA fragment having Sau3A sticky ends.
The plasmid to be used for the cloning experiment has the following MCS
EcoR1, BamH1, Xba1, HindIII

*Describe how you would do the cloning experiment.

Restriction enzyme recognition sites
EcoR1 = GAATTC (the enzyme cuts after the G)
BamH1 = GGATTC (the enzyme cust after the first G)
Xba1 = TCTAGA (the enzyme cuts after the first T)
HindIII = AAGCTT (the enzyme cuts after the first A)
Sau3A = GATC (the enzyme cuts before the G)

#2 bob1


    Thelymitra pulchella

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Posted 01 October 2012 - 11:39 PM

Your BamH1 is wrong - should be GGATCC - which will help you in your cloning (think which ends will be compatible with Sau3a).

As this is homework, I'm not giving a full answer, you need to provide a bit more input so that we can help you learn rather than just having us supply the answer.

#3 susnata



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Posted 26 July 2013 - 01:45 AM

i think you already got the answer... it's called "compatible cohesive ends"

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