Degradation time of 3' A overhang in TA cloning

[Andrew Harrison]

Have you thought about reamplifying your faint band by spiking it on a 
gel with a syringe needle, and then reamplifying from the "spiked" band? 
We find this works very well and might save you having to gel purify 
bands etc.(vortex the syringe needle in about 20ul of water and use 10 ul 
for the next PCR)
We have also used Promega "Magic Clean-up" columns to clean up PCR 
products before cloning (although we later found this to be unnecessary, 
and didn't have any problems with the TA vector which seems to be very 
reliable.

Hope this is of help,
Hilary J. Rogers