By Anonymous on Thursday, May 30, 2002 - 02:25 am:
Is there any conventional LC column (C8 or C18 type) for the analysis of DNA molecules (about 10.000pb. Any information related to column type (particle size, pore size)as well as mobile phase composition will be helpfull. I have seen several applications described by Waters, but they are only applicable to small oligos.
Thanks in advance
Samir
By Anonymous on Monday, August 26, 2002 - 11:45 am:
Does it have to be RPC?
TosoHaas and Amersham Biosciences have non-porous bead, anion exchange columns suitable for macromolecules.
Indeed, TosoHaas have an application note on the purification of PCR products using their DNA NPR column.
By Jim Thayer on Monday, August 26, 2002 - 03:29 pm:
DNA Analysis by RPLC:
Ion-Pairing methods for dsDNA work up to 1K-2K bps (see Huber et.al, NAR 21(5):1061 ff, 1993.
These phases are either nonporous (~2µm dp) or have at least 700Å pores for ~3.5µm dp phases. All appear to be C18.
Anion Exchange phases for dsDNA are typically non-porous, and may be very small (~ 2.5µm dp, DNA-NPR: TosoHaas) or relativeley large (~13 µm dp, DNA Pac PA100: Dionex). These phases require very careful control of gradient slope to achieve separations of > 2K bp. Use of systems with curved gradient support should help.