solubility of dna
Posted 27 June 2004 - 04:10 AM
i´m looking for the best method to dissolve dna-pellets,after it is extracted. the pellet is not completely dissolved in TE ph 8,0 and therefor the pcr shows not the results it should be. do you the best way to dissolve the dna-pellet??please tell me!!thank u..
Posted 27 June 2004 - 10:14 AM
Posted 27 June 2004 - 04:35 PM
I agree with hula, it could be something other than dna that you've isolated. Otherwise could be a number of things:
1) Did you have an ethanol wash as the last step of your protocol? If the ethanol wasn't dried off properly your nucleic acid will not dissolve. I normally dry my samples with a pipete tip (capillary action, upside down) and then leave them for a while with lids open (cover with tissue or something) on bench or ona heating block (not too hot). With the samples you alredy have you can do an ethanol precipitation and redissolve
2) Your dna might be very concentrated and so reached the extent that it'll dissolve, what's the concentration? Try adding more buffer.
3) Leave your dna at 4C overnight to allow to dissolve, genomic dna takes a while to dissolve
Hope this helps!
Posted 27 June 2004 - 04:56 PM
Posted 29 June 2004 - 05:49 PM
Also, sometime with DNA dissolved in TE one has to slightly increase MgCl2 concentration... usually 0.25 mM is enough... depends on volume of DNA you are adding though...