Yeast tRNA as carrier in the isolation of microscale RNA for global amplification and expression profiling.
The characterization of global gene expression patterns of microscale samples is important in many areas of biological and clinical research. The choice of carrier is critical for the efficient isolation and successful amplification of RNA at the nanogram level. Here we show that recovery of nanograms of RNA is significantly higher when carrier linear polyacrylamide is supplemented with carrier tRNA. Reverse transcription and in vitro transcription reactions remain efficient and specific in the presence of carrier tRNA. Finally, comparison of GeneChip array hybridization patterns demonstrates that the presence of carrier tRNA does not cause detectable distortion in global amplification. Taken together, tRNA is a superior carrier for the isolation and global amplification of microscale RNA.
Wang
Q Tian
QT
Department of Biochemistry, Stanford University School of Medicine, CA 94305-5307, USA. qwa205@stanford.edu
Xiao
Wenzhong
W
Mindrinos
Michael
M
Davis
Ronald W
RW
D002478:Cells, Cultured; D004847:Epithelial Cells; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D012331:RNA, Fungal / Q000302:isolation & purification* / Q000378:metabolism; D012343:RNA, Transfer / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics
D012331:RNA, Fungal; D012343:RNA, Transfer