agarose gel of ligated DNA doesn't show any bands
Posted 08 June 2010 - 08:45 AM
Posted 08 June 2010 - 10:10 AM
How long do you incubate the reactions?
What buffer do you use for elution after gel extraction?
Maybe some of your ingredients for the reactions is contaminated with nucleases?
Check if your sample is also degraded within the incubation time without adding insert, vector, ligase, ligase buffer, water and so on.
Posted 09 June 2010 - 04:42 AM
As a consequence, You put very low amount of DNA in the ligation reaction and therefore, You're not able to see it on gel.
I'd recommend to run a small amount of eluted DNA after gel extraction to make sure it works well.