Quantitative Mammalian Transcriptomics using Designed Primer-based Amplification

ABSTRACT: We describe a novel quantitative cDNA expression profiling strategy, involving amplification of the majority of mouse transcriptome using a defined set of 44 heptamer primers. The amplification protocol allows for efficient amplification from as low as 50pg of mRNA and did not alter the expression of the transcripts even with 200 fold dilution of the minimum requirement of the starting material (10ng of mRNA) for standard RNA-seq protocols. We implemented our methodology on embryological lineage segregation, achieved by graded activation of Activin A/TGFβ signaling in mouse embryonic stem cells (mESCs). The fold changes in transcript expression were in excellent agreement with quantitative RT-PCR and we observed a dynamic range spanning more than five orders of magnitude in RNA concentrati

ORGANISM(S): Mus musculus

SUBMITTER: vipul bhargava 

PROVIDER: E-GEOD-45474 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress