cbil.upenn.edu:RAD.Protocol:925:Protocol - cbil.upenn.edu:RAD.Protocol:925:Protocol

Type
nucleic acid labeling protocol
Description
See pages 2.1.10-2.1.17 of the GeneChip Expression Analysis Technical Manual for all details. First-strand cDNA is synthesized from (5.0 to 20.0 ug) high-quality total RNA, using the GeneChip T7-Oligo(dT) Promoter Primer Kit or random primers provided with the SuperScript Choice Kit. Second-strand synthesis is carried out in: 1X second-strand reaction buffer, 200 uM each dNTP mix, 10 U E. coli DNA ligase, 40 U E. coli DNA polymerase I, 2 U E. coli RNase H in a final volume of 150 uL. The double-stranded cDNA is cleaned-up using the components supplied with the GeneChip Sample Cleanup Module. Enzo BioArray HighYield RNA Transcript Labeling Kit (Affymetrix, P/N 900182) is used to synthesize biotin-labeled cRNA target. The latter is cleaned up using the components supplied with the GeneChip Sample Cleanup Module.
Links
All 12 experiments using protocol cbil.upenn.edu:RAD.Protocol:925:Protocol