Hello Everybody,
We are working with faecal samples to research the gut microbiota with DGGE. We want to know what is the Synthesis Scale of the primers that you use in the first PCR?
Thank you very much.
it depends on whether you are using a GC clamp on the first round of pcr. If yes then ther length of the primer plus clamp will be about 50 bases so you will need purification of the clamped oligo so this will probably not be available from your oligo supplier so the lowest scale which will give you an HPLC purification option. If you are amplifying with short primers then a re amplification with a clamped oligo the desalted pcr primers at the lowest scale and then the clamped ologo at the lowest scale that will allow HPLC purification. If cost is a problem for purified longmers you might want to look into binding the pcr product together with psoralen which had much the same effect as a GC clamp but only need a couple of A bases at the 5. end of the primer which were bound together using UV light.
One primer containing the GC clamp can serve the purpose(in single PCR only)
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology