cbil.upenn.edu:RAD.Protocol:405:Protocol - cbil.upenn.edu:RAD.Protocol:405:Protocol
mRNA (~1 ng) was amplified from 100 ng total RNA (equivalent to ~10,000 endothelial cells) using the MessageAmp aRNA Kit (Ambion, Austin, TX). Poly(A) RNA was reverse transcribed using an oligo(dT) primer containing a T7 RNA polymerase promoter sequence. RNase H treatment cleaved the mRNA into small fragments that served as primers during second-strand synthesis, resulting in a double-stranded cDNA template for T7-mediated linear amplification by in vitro transcription. Typically 4-5 ug aRNA were produced from one round of amplification (a 4,000-5,000 fold amplification).
|All experiments using protocol cbil.upenn.edu:RAD.Protocol:405:Protocol: (E-CBIL-17, E-CBIL-3, E-CBIL-32, E-CBIL-41, E-CBIL-42, E-CBIL-43, E-CBIL-47)|