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Old 03-09-2012
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DNA Cloning and recombinant plasmid

cDNA (human growth hormone gene) is cloned into recombinant plasmid and transferred into bacterial gene to produce HGH.
During cloning same restrictino enzyme sequence is emplyed at BOTH 5' and 3' of gene to insert the coding sequence after promotor sequence.

how many percent do cloned bact share and why?
ans: 50%
why not 100%??
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Old 03-09-2012
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This is because with the same restriction site on both ends of the inserts (some people are comfortable calling it overhangs, the sticky ends,etc), you have an equal chance of getting the orientation of the insert to be correct (50%) and wrong (50%)

if you used two diff restriction enzymes, you can be positive that the 5' end of the inset will join with the 3' end of the vector, giving you a correct oriented insert.

hope this helps!
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The above post was thanked by:
venky2600 (08-18-2012)


Genetics-, Microbiology-

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