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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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gel purification
Posted by: sandesh regmi (IP Hidden, Unregistered user, )
Date: January 26, 2005 12:32PM
I have encountered a weired problem while purifying my vector for ligation. I double gigest my vector and ran in the gel. It looked pretty nice, well separrated two bands. I excised the band from the gel ( 4KB), purified by using QIAkit. I again ran an aliquot in the gel just to make sure that I have right things. But unfortunately I have two bands one has expected size and the pther has reduced size. I heat inactivated my reaction. Everytime I got reduced bands, what would be the reason? If anyone has a solution I would be veryy happy. Thanks
sandesh regmi
Re: gel purification
Posted by: Jula (IP Hidden, New member, 4)
Date: January 26, 2005 01:11PM
Hello,
Have you run the undigested vector next to it as a control? Maybe you have to dephosphorylate it i.e with SAP, so the ends won't stick together again - I guess that your second band might be the circular plasmid? Good luck.
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