Strain: C57BL6 BLK.CL4 total RNA (MoBC.29xx) 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 MoBC.29xx.sig21 Cell type BLK.CL4 Source NHGRI RNA isolation LYNX mRNA QC passed cDNA library: Library DpnII restriction - (signature cloning using MmeI) Sequence length 20 bp MPSS: runs MoBC.29xx_sig21.4788W.a-20 12/17/2003 588761 13368 MoBC.29xx_sig21.4788T.b-20 12/9/2003 451422 11263 MoBC.29xx_sig21.4788T.a-20 12/9/2003 429623 11932 MoBC.29xx_sig21.4788W.b-20 12/9/2003 531391 11722 Run group: Total Beads successfully sequenced - 2001197 Processed Signatures - 15689 Transcripts 3'-most (Canonical) Signatures - 10081 Lot batch =
