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A: Digital gene expression / SAGE experiment. B: Most frequent TagDust filtered sequences. TCGTATGCCGTCTTCTGCTTG 35191 GTATGCCGTCTTCTGCTT 4733 AA- TCGTATGCCGTCTTCT 3109 TCGGAC - TCGTATGCCGTC 2963 G- TCGTATGCCGTCTTCTG 2875 TCGGACTGTAGAA TCGTA
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A: Digital gene expression / SAGE experiment B: Most frequent TagDust filtered sequences TCGTATGCCGTCTTCTGCTTG 35191 GTATGCCGTCTTCTGCTT 4733 AA-TCGTATGCCGTCTTCT 3109 TCGGAC-TCGTATGCCGTC 2963 G-TCGTATGCCGTCTTCTG 2875 TCGGACTGTAGAATCGTA 2818 ATGGC-TCGTATGCCGTCT 2339 AGGAG-TCGTATGCCGTCT 2307 TC-TCGTATGCCGTCTTCT 2108 TCGGACTGTAGAACTCTT 1936 A-TCGTATGCCGTCTTCTG 1886 AGGAGT-TCGTATGCCGTC 1880 CGTATGCCGTCTTCTGCT 1667 GTATGCCGTCTTCTTCTT 1527 AG-TCGTATGCCGTCTTCT 1509 CCAG-TCGTATGCCGTCTT 1366 GTGA-TCGTATGCCGTCTT 1238 GTG-TCGTATGCCGTCTTC 1209 TCGGACTGTAGA-TCGTAT TCGGACTGTAGAACTCTGAAC 1907 GTTTCAGGAGTTTATTTT ACAGGTTCAGAGTTCTACAGTCCGACATG 1268 GCCACCCTCTACAG-CCGA mRNA enrichment with polyT beads GEX Adapter 2.1 NlaIIIrecognition site Poly-A-Tail Bead 5' 3' NNN...NNNCATGXXXXXXXXXXXXXXXNNN...NNNAAAAAAAAAAA 17nt sequence mRNA NNN...NNNGTACXXXXXXXXXXXXXXXNNN...NNNTTTTTTTTTTT cDNA 1st and 2nd strand synthesis Bead NNN...NNNCATGXXXXXXXXXXXXXXXNNN...NNNAAAAAAAAAAA NNN...NNNGTACXXXXXXXXXXXXXXXNNN...NNNTTTTTTTTTTT 1st Digestion NlaIII Bead NNN...NNNCATGXXXXXXXXXXXXXXXNNN...NNNAAAAAAAAAAA NNN...NNNGTACXXXXXXXXXXXXXXXNNN...NNNTTTTTTTTTTT Ligation GEX adapter 1 Bead ...TCTACAGTCCGACATGXXXXXXXXXXXXXXXNNN...NNNAAAAAAAAAAA ...AGATGTCAGGCTGTACXXXXXXXXXXXXXXXNNN...NNNTTTTTTTTTTT 2nd Digestion GEX Adapter 1.2 MmeIrecognition site Bead ...TCTACAGTCCGACATGXXXXXXXXXXXXXXXNNN...NNNAAAAAAAAAAA Matches human NlaIII restriction site chr20:+1868416-1868436 ...AGATGTCAGGCTGTACXXXXXXXXXXXXXXXNNN...NNNTTTTTTTTTTT Ligation GEX adapter 2 GEX Adapter 1.1 Sequencing primer site ...CAGAGTTCTACAGTCCGACATGXXXXXXXXXXXXXXXTCGTATGCCGTCTTCTGCTTG CAAGTCTCAAGATGTCAGGCTGTACXXXXXXXXXXXXXNNAGCATACGGCAGAAGACGAAC