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bio25

Member Since 22 Jan 2013
Offline Last Active Jan 25 2013 06:26 AM
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Topics I've Started

Yeast two hybrid

25 January 2013 - 06:14 AM

Hi everyone,
I am presently performing Y2H. I am earnestly looking for the Y187 strains. If someone can help me, shall remain ever grateful.



Thanks in advance.

SalI and double digestion.

22 January 2013 - 04:59 PM

Dear members,

I have performed restriction digestion with SalI and SmaI in a sequential mode. While cutting with SalI itself the vector and the insert got separated. (when it can only happen with double digestion). So, to check it further, when I purified the gel and cut the insert and further digested with SmaI, I can see a faint band of my gene only with the correct band size as per the case of SalI. This apparently seems to be correct as SmaI only has the gene to be excised, no vectors to separate my gene from. But, my query is why the case happened like this with SalI? why did it separate my gene of interest at the single cut? is there any specific property for the same? or I am missing something on the way.. Please if anyone can guide me...

Note: SalI and SmaI are the enzymes that do not cut the insert. So, I selected the same.

Keenly looking for some guidance at the earliest...

Thanks in advance..

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