SAMPLE PREPARATION METHODS

摘要

The present invention provides whole blood nucleic acid extraction methods, compositions, and kits, as well as nested isothermal amplification methods, compositions, and kits. In certain embodiments, these methods are applied to detecting Lyme disease, including in patients without classic erythema migrans skin lesions.

主权项

1. A method of nucleic acid extraction comprising:
a) contacting a sample whole blood with beads, a proteinase, and an anionic surfactant to generate a treated sample; b) homogenizing said treated sample to generate a cell lysate; c) centrifuging said cell lysate comprising a supernatant; d) separating said supernatant from said cell lysate; e) adding magnetic particles and lysis buffer to said supernatant to generate a magnetic-particle sample, wherein said magnetic particles are configured to bind nucleic acid molecules; f) washing said magnetic-particle sample with a wash buffer; g) treating said magnetic-particle sample in order to generate a dried magnetic bead sample; and h) treating said dried magnetic bead sample with an elution buffer such that a purified nucleic acid sample is generated that comprises purified nucleic acid.