Multiple bands from purified PCR product
Posted 08 April 2011 - 05:09 AM
I am studying alternative splicing of a certain gene under different conditions.
So I did some PCRs and saw multiple products on gel. I then purified some of these products from the gel and re-did the PCR using the same primers for sequencing. Strangely, I saw that after the 2nd PCR of one of the products (indicated in red on the left gel), I obtained two (maybe even three) products. The size of the smallest product in the middle gel corresponds to the faint band on the left gel. So I purified these two products, again re-did the PCR and still I obtain multiple bands in the reaction with the largest product.
Can anybody explain this?
Posted 08 April 2011 - 05:58 AM
Posted 08 April 2011 - 06:12 AM
Anyway, even with non-specific primers you shouldn't get a 200 bp product from a purified 600 bp template.
Posted 25 April 2011 - 01:27 PM