Whole blood sample preparation for polymerase chain reaction usi

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage

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435 912, 435 6, 536 243, 536 2432, 424534, C12Q 170, C12P 1934, C07H 2104

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active

057028842

ABSTRACT:
Leucocytes can be quickly and selectively separated from erythrocytes by subjecting a whole blood sample to a series of steps including lysing the erythrocytes and washing the remaining leucocytes with a solution containing ammonium chloride and a carboxylic acid or a metal carboxylate. The resulting white blood cells can be readily lysed and subjected to polymerase chain reaction to amplify and detect a target nucleic acid. The test kit useful in practicing the amplification method includes a labeled primer, a PCR reagent and a reagent mixture containing ammonium chloride and a carboxylic acid or a metal carboxylate for sample preparation.

REFERENCES:
patent: 4407942 (1983-10-01), Birnboim
patent: 4683195 (1987-07-01), Mullis
patent: 5300635 (1994-04-01), Macfarlane

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