Hello everyone I look for a purification protocol of DNA primer by HPLC after being synthesized by a oligosynthetiseur
Thank you
Hello Mouna
in deed there is no universal protocol for primer purification , because this depend on the type of the column that you have , and to be more clear each column type has its conditions for the best performance .
and you have to be sure that you have the right column that is design specifically for macromolecules which always have a pore size more than 200 angstrom ,
so tell me what column you have and i will try to help you .
note : you should have preparative or semi-preparative HPLC for this purpose .
Regards
hello Mouna
well in this case i can suggest the following protocol
if you use 4.6 mm ID column the flow rate will be 2ml/min ( you should use an other flow rate if you have an other column ID )
the mobile phase is 10% acetonitrile and 90% of 0.1M acetate at PH 7.5
the monitoring on 260 nm
50uL or lesser sample size . ( this depend on the column ID , )
good luck
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