I am ligating a promoter (pCadA) and RBS to make a composite part. This will then be inserted into a plasmid backbone. When i ran PCR products on Gel against a 50 base pair ladder. The results were confusing, the 12 bp part band appeared against band more than 50 bp.
Regards.
Hi Shaheer,
if the fragments that you are trying to ligate are so small, it seems that the easiest way would be to order your entire DNA fragment as an oligo. Then you can insert it into your vector.
Best of luck!
That was the first thing that came to my mind as well, but we are short on time. We have got to submit our parts to the registry by 27th of October.
I'm on a team competing in iGEM!
Do the small fragments have overlapping parts such as compatible sticky ends?
If so, your best bet is to heat denature them and anneal by slow cooling back to room temperature. This will yield composite fragments that you can bond with ligase. Then just digest to get the sticky ends for the vector and ligate to the vector.
Hopefully this is possible with the configuration of the fragments that you have...
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