separation of dna fragments in agarose gel electrophoresis
Posted 01 June 2009 - 09:42 AM
im trying 2 elute the insert fragment ffrom gel for cloning purpose.the problem is that the insert fragment is around 2 kb and the vector backbone is around 2.1kb.
im trying to run these fragments on 1% agarose gel but the separation is nt much as im nt able 2 distinguish.Is 0.8% agarose gel more suitable than increasing the agarose concentration of the gel.how can i get my insert separated?
thanks in advance
Posted 01 June 2009 - 10:45 AM
Posted 01 June 2009 - 01:25 PM
If you know the sequence of your vector and of your insert, your best be is to do as phage434 suggests...
Posted 01 June 2009 - 03:03 PM
Posted 01 June 2009 - 07:26 PM
Posted 01 June 2009 - 08:18 PM
its nt possible 2 cut it any where else coz i cloned my insert into the plasmid with promoter ,terminator and now i want to take my insert out of the vector along with promoter and terminator.i have EcoRV to do so after that i get vector and desired insert fragment.
i think i should try lower concentration of agarose gel
Posted 02 June 2009 - 04:28 AM
Posted 02 June 2009 - 06:43 AM
can polyacrylamide be used? normal elution kits that elute dna from agrose ,can they do that for polyacrltamide also??