Other

Ge Healthcare Bio-Sciences Corp.

Pharmacia Biotech Inc.

R Scott Duthie

John Richard Nelson

Gregory Andrew Grossmann

Charles K Brush

Anuradha Sekher

William H Hildebrand

Gregory Andrew Grossman

Rohini Dhulipala

Mary Ellexson

Pierre Chretien

Lawrence J Burazin

Mark E Freeman

Eugene P Stirchak

Methods of amplification, purification and detection of nucleic acid sequences especially RNA are described. One aspect of the method involves the hybridisation and subsequent ligation of a nucleic acid structure to the nucleic acid sequence desired to be manipulated. The methods require that the nucleic acid structure comprises a double stranded region and a single stranded region. The single stranded region is complementary to the RNA sequence of interest. The double stranded region may also contain additional functionalities which are then used subsequently in the method.

Ligation-based RNA amplification

There is provided a method for directly typing or sequencing HLA-A, -B, or -C alleles from a tissue sample wherein exons 2 and 3 of the HLA-A, -B, or -C alleles from the sample are amplified together in a locus specific manner and then separated out and individually amplified in a locus specific manner. After the two amplifications, the amplified exons are directly sequenced, the sequences are recombined, and a comparison is made between the derived HLA allele sequence and an HLA allele database, thereby giving an exact HLA-A, -B, or -C type for the sample being tested.

Class I sequence based typing of HLA-A, -B, and -C alleles by direct DNA sequencing

A preparation of a labelled nucleotide comprising at least one compound having a Mg.sup.2+ association constant between 1.times.10.sup.-11 to 1.times.10.sup.-2, inclusive. The compound is preferably selected from the group consisting of citrate, isocitrate, phosphate, EGTA, EDTA, and CDTA. The concentration of the compound is preferably at least 5 mM.

Composition for stabilization of labelled nucleoside triphosphates and

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R Scott Duthie