Long PCR stuck in the wells :-( %8) warm samples before loading

L. Sumoy Sumoy at mbcg.uchc.edu
Sat May 28 12:49:20 EST 1994


In article <2s5pfg$r3t at gazette.bcm.tmc.edu>
hdang at cns.neusc.bcm.tmc.edu (Hong Dang) writes:

... heavy EtBr band stuck in the well. 

Warm samples at 65 degrees C for 5 minutes and chill in ice water
before loading on the agarose gel. We do this for genomic DNA
digestions for Southerns to prevent DNA from staying in the well or
runing abnormally. Sticking in well could be due to precipitation or to
annealing between sticky ends of very large fragments of DNA, or to
high concentration of protein contaminants. Heating samples may help
dissolving and preventing aggregation.
Whatever it does it seems to solve the problem.
Good luck.
Lauro. 



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