Transcription profiling of mouse tissue samples to assess an RNA amplification and labelling methodology

ABSTRACT: We have developed a total RNA amplification and labeling strategy for use with Affymetrix GeneChips. Our protocol, which we denote BIIB, employs two rounds of linear T7 amplification followed by Klenow labeling to generate a biotinylated cDNA. In benchmarking studies using a titration of mouse universal total RNA, BIIB outperformed commercially available kits in terms of sensitivity, accuracy, and amplified target length, while providing equivalent results for technical reproducibility. BIIB maintained 50 and 44% present calls from 100 and 50 pg of total RNA, respectively. Inter- and intrasample precision studies indicated that BIIB produces an unbiased and complete expression profile within a range of 5 ng to 50 pg of starting total RNA. From a panel of spiked exogenous transcripts, we es

ORGANISM(S): Mus musculus

SUBMITTER: Jeff Shearstone 

PROVIDER: E-GEOD-5130 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress