Method for conducting sequential nucleic acid hybridization step

Details Method for conducting sequential nucleic acid hybridization step Method for conducting sequential nucleic acid hybridization step

1994-06-06

1998-06-30

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 911, 435 912, 536 2032, 536 2433, C07H 2102, C07H 2104, C12P 1934, C12Q 168

Patent

active

057732132

ABSTRACT:
A method for conducting sequential nucleic acid hybridization steps is described, whereby the ability of earlier-used primers or probes to participate in subsequent hybridization steps can be minimized, even though the differences between primer lengths are relatively small. It also relates to a rapid and quantitative method for the sequential synthesis of polynucleotide sequences by using a plurality of oligonucleotide primers, with the earlier utilized primers causing a minimum of interference with the subsequent primed synthesis reactions, yet without the need for intermediate purification steps. One preferred embodiment described is a method for differential display reverse-transcription polymerase chain reaction (DDRT-PCR), wherein complementary DNAs (cDNAs) are first synthesized using oligo-dT-primed reverse transcription (RT), and selected subsets of said cDNAs are then amplified using a second primer in a polymerase chain reaction (PCR), with a minimum degree of background being caused in the PCR step by residual amounts of the oligo-dT primer.

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