Digital transcriptome profiling using selective hexamer priming for cDNA synthesis.
We developed a procedure for the preparation of whole transcriptome cDNA libraries depleted of ribosomal RNA from only 1 microg of total RNA. The method relies on a collection of short, computationally selected oligonucleotides, called 'not-so-random' (NSR) primers, to obtain full-length, strand-specific representation of nonribosomal RNA transcripts. In this study we validated the technique by profiling human whole brain and universal human reference RNA using ultra-high-throughput sequencing.
Armour
Christopher D
CD
Department of Molecular Informatics, Rosetta Inpharmatics LLC, a wholly owned subsidiary of Merck and Co., Inc., Seattle, Washington, USA. christopher_armour@merck.com
Castle
John C
JC
Chen
Ronghua
R
Babak
Tomas
T
Loerch
Patrick
P
Jackson
Stuart
S
Shah
Jyoti K
JK
Dey
John
J
Rohl
Carol A
CA
Johnson
Jason M
JM
Raymond
Christopher K
CK
D001921:Brain / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D006801:Humans; D012313:RNA / Q000235:genetics / Q000378:metabolism
D018076:DNA, Complementary; C034477:RNA primers; D012313:RNA