Strain: C57BL6 Brain:Midbrain, Male total RNA (MoBr.MaMb) received from National Human Genome Research Institute was analyzed through Bioanalyzer and passed our quality control test. The mRNA was processed according to the MPSS protocol as outlined in the previous publications (Brenner, S., et al. (2000) Proc Natl Acad Sci U S A 97(4): 1665-1670 and Brenner, S. et al., Nat. Biotechnol 18(6): 630-634). Briefly, the mRNA was reverse transcribed and the cDNA was digested with Dpn II. The 20 bases adjacent to the 3? most Dpn II site was cloned into a Megaclone vector. The resulting library was amplified and loaded onto microbeads. About 1.6 million microbeads were loaded into each flow cell and the signature sequences were determined by a series of enzymatic reactions as outlined in the above publications. The abundance for each signature was converted to transcripts per million (tpm) for the purpose of comparisons between samples. Cells/tissue: Library MoBr.MaMb.sig21 Cell type Brain:Midbrain, Male Source NHGRI RNA isolation LYNX mRNA QC passed cDNA library: Library DpnII restriction - (signature cloning using MmeI) Sequence length 20 bp MPSS: runs MoBr.MaMb_sig21.5169F.c-20 9/30/2004 350976 11217 QC Passed MoBr.MaMb_sig21.5169W.c-20 9/30/2004 619979 18100 QC Passed MoBr.MaMb_sig21.5169W.a-20 9/14/2004 551415 14529 QC Passed MoBr.MaMb_sig21.5169F.b-20 9/13/2004 592802 17444 QC Passed Run group: Total Beads successfully sequenced - 2115172 Processed Signatures - 22737
