Low yield PCR product after gel purification
Posted 13 March 2013 - 07:12 PM
I need everyone help and suggestion. I have try to purified PCR product from gel purification kit. I've check PCR product by electrophoresis I found that I got thick band of my target product but after I purified I got very low yield ( 9 ng/ul). So I try another way, by precipitate PCR product first and measure conc. got 1.4 ug/ul. Then, gel purification with kits. Unfortunately, I still got low yeild (9-10 ng/ul). I have more reviewed. I found another way. that's using enzyme agarase for digest gel directly not need use column purification that maybe couse low yield. Everyone, How do you think about this way and have another techniques for gel purification with high yield , I just need 100ng for go next step in my study. Thanks all for reading and suggestion. I'm look forward you helping.
Posted 13 March 2013 - 07:50 PM
Are you positive your dilution concentrations are correct? What is the final volume of your DNA elute? You do not have enough for 100ng?
Posted 14 March 2013 - 02:09 AM
Posted 14 March 2013 - 04:16 AM
talent does what it can
genius does what it must
i used to do what i got paid to do
Posted 14 March 2013 - 11:49 AM
Also note 4 x30 = 120 ng... You could try concentrating the DNA by evaporation.
Posted 14 March 2013 - 05:14 PM
Posted 20 March 2013 - 06:37 AM
Posted 26 May 2013 - 07:23 PM
I had the same problem with Quigen kit and after doing so many trials I managed to increase my product concentration upto ~100ng/uL.
Good luck !!!