AMPLIFICATION AND ANALYSIS OF WHOLE GENOME AND WHOLE TRANSCRIPTOME LIBRARIES GENERATED BY A DNA POLYMERIZATION PROCESS

摘要

The present invention regards a variety of methods and compositions for whole genome amplification and whole transcriptome amplification. In a particular aspect of the present invention, there is a method of amplifying a genome comprising a library generation step followed by a library amplification step. In specific embodiments, the library generating step utilizes specific primer mixtures and a DNA polymerase, wherein the specific primer mixtures are designed to eliminate ability to self-hybridize and/or hybridize to other primers within a mixture but efficiently and frequently prime nucleic acid templates.

主权项

1. A method of preparing a plurality of nucleic acid molecules having known constant region at each end, comprising:
a) obtaining a sample comprising nucleic acid molecules; b) subjecting said nucleic acid molecules to a population of primers to form a nucleic acid molecule/primer mixture, wherein the primers of the population have a nucleotide sequence that is substantially non-self-complementary and substantially non-complementary to other primers in the population, wherein the primers are composed of a constant region and a variable region, the constant region being positioned 5′ to the variable region, wherein primers in the population comprise non-complementary and non-self-complementary nucleotides selected from guanines and adenines; cytosines and thymidines/uridines; adenines and cytosines; or guanines and thymidines/uridines in a relative proportion effective to render the polynucleotides substantially incapable of at least one of the following: self-hybridization; self-priming; hybridization to another polynucleotide in the plurality; or initiation of a polymerization reaction in the plurality; and c) subjecting said nucleic acid molecule/primer mixture to a polymerase under conditions to generate the plurality of molecules including the known constant region at each end.