what's the proper amout of 16SrDNA should I transfer to DGGE - (Jan/19/2005 )
After PCR of 16SrDNA,I subject 16SrDNA to DGGE,but I don't know what's
the proper amount of 16SrDNA should I transfer to DGGE,I mean if I add
more than the proper amount,It would be wasteful;if I add less than the
proper amount,some bands couldn't be detected by DGGE.
Could you please give me some suggestion about it?
I have the same question as you.Do you solve it yet?
In general, for a complex community of microorganisms, one could add 300-500 ng of PCR product. However, this can vary with the complexity of the community and the sensitivity of your stain.
Just to let you know, I have put together a website for DGGE which incorporates the experience of many DGGE researchers. I hope this will help: http://ddgehelp.blogspot.com/