Strain: C57BL6 Large intestine, Male total RNA (MoLI.Male) 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 MoLI.Male.sig21 Cell type Large intestine, Male Source NHGRI RNA isolation LYNX mRNA QC passed cDNA library: Library DpnII restriction - (signature cloning using MmeI) Sequence length 20 bp MPSS: runs MoLI.Male_sig21.5193F.d-20 11/4/2004 429787 4715 QC Passed MoLI.Male_sig21.5193F.c-20 10/27/2004 322659 3331 QC Passed MoLI.Male_sig21.5193W.b-20 9/28/2004 386294 4631 QC Passed MoLI.Male_sig21.5193W.a-20 9/28/2004 405911 4330 QC Passed MoLI.Male_sig21.5193F.b-20 9/27/2004 390518 5880 QC Passed Run group: Total Beads successfully sequenced - 1935169 Processed Signatures - 7488
