0
Im having extreme trouble attempting to generate a library with a gene length of 900bp.
The problem being, we are wanting to randomise many (1-10) regions each of approx. 20-30bp, without any templeate at all. Therefore, this whole procedure is a synthetic gene repetoire made entirely from primers.
We get a lot of smearing and often only smearing and no bands. We have played around with the temps, amount of pcr template, Mg concentration etc but are having lots of trouble.
Usually we get 2 bands, one the correct size and one 50bp higher.
I have spent a long time on this and i cannot see a surefire way to get reproducible bands.
Please can someone help!
valetudogirl
