Smear after second PCR
Posted 12 August 2004 - 11:26 PM
Posted 12 August 2004 - 11:32 PM
Posted 16 August 2004 - 06:08 AM
Hi I read somewhere that diluting your starting product could help.
As the template is a PCRed product, the concentration is still quite high compared to the amount of template you normally add when doing PCR.
Therefore, probably, a dilution of 1:10 of the PCR product followed by loading 1ul as template would do the trick. Also, lesser amount of cycles should be used -to prevent mutation to products as well as resulting in smear like result.
Hope the above helps. Have personally not tried it but I read it somewhere in this or another forum.