ISOLATION AND DETECTION OF DNA FROM PLASMA

摘要

Provided herein is technology relating to the amplification-based detection of bisulfite-treated DNAs and particularly, but not exclusively, to methods and compositions for multiplex amplification of low-level sample DNA prior to further characterization of the sample DNA. The technology further provides methods for isolating DNA from blood or blood product samples, e.g., plasma samples.

主权项

1. A method of processing a plasma sample, the method comprising:
a) combining a plasma sample with
i) protease;ii) a first lysis reagent, said first lysis reagent comprising
guanidine thiocyanate,non-ionic detergent; to form a mixture wherein proteins are digested by said protease; b) to the mixture of step a) adding
iii) silica particles, andiv) a second lysis reagent, said second lysis reagent comprising:
guanidine thiocyanate;non-ionic detergent; andisopropyl alcohol, under conditions wherein DNA is bound to said silica particles; c) separating silica particles with bound DNA from the mixture of b); d) to the separated silica particles with bound DNA adding a first wash solution, said first wash solution comprising guanidine hydrochloride or guanidine thiocyanate and ethyl alcohol; e) separating the silica particles with bound DNA from said first wash solution; f) to the separated silica particles with bound DNA adding a second wash solution, said second wash solution comprising a buffer and ethyl alcohol; g) separating the washed silica particles with bound DNA from said second wash solution; h) eluting DNA from said washed silica particles with bound DNA to produce an isolated DNA sample.