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Recombinant DNA

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#1 biobimbo



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Posted 04 March 2005 - 02:57 PM

I am enrolled in a biotech lab 1 course at a local college. For the past 2 weeks we have been conducting experiments on cloning and selection of dna fragments. We used a pUC19 vector and the human follistatin fragment. After all the steps of gel isolating, transformation, digesting and ligation my final gel analysis revealed 2 fragments. The professor said I had a 250bp fragment and another fragment that was a "double insertion".
Why did this occur? Was it during ligation with the ligase where 2 copies of my hfs recombined together? Did it occur afer the digestion with Pst1? I don't know how to make a linear map because I can't figure out what happened. I would appreciate your input!

#2 tfitzwater



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Posted 07 March 2005 - 04:31 PM

You need to tell us what restriction enzymes you used to cut the insert and vector so we know what was supposed to happen.

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