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DNA preparation and MLVA protocol

par Christine Pourcel - 21 août 2009

DNA preparation For PCR, DNA can be purified using commercial kits or alternatively pure DNA is prepared using CTAB and Phenol/chloroform extraction. The DNA is kept frozen at -20 °C.

MLVA

A first study by Onteniente et al. in 2003 described 6 VNTRs and showed that MLVA genotyping was as efficient and informative as ribotyping and PFGE.

In order to further increase the efficiency of MLVA, we made use of the sequenced genome data for reference strain PA14, to identify new VNTRs.

The present MLVA scheme includes 13 minisatellites ( repetition longer than 9bp) and 2 microsatellites.