Comment: early pupae
TGTGAACTGAGACTCGGAGATACAGCGGAAAGGATCTAGAAGATACGCGTAGTTTGGCCCACCAAAACAACTACTGACCG AACGGATTGTTGTGGCCCAGTGAAGTTATCGTTGAAATTGGAAATTGATCAAATGAATAAATGTATCCGCGAGATACGCC CAGTGAAGTTTTAATTTTCTAGCCGGAGAGAAAAATCAAGTGTGAAAAGCAAAAAGGAAAAACGAAAAACGAAAAACCGC CCCCACAACAATAACAAAAACAACCGGTACGCCACCCCCCGAAAAATTGACGTCGCCGTCGCCCTCGTCGCAATCGTTTG GAAATAAATCGTGTTTTTGCATTTGTTGTTTTTTGTTGTTTATACGCCAAGTTATTTACGTCGTCGCAAAGCAAAAAGAA AAAATTGAATAAATAACAACAAAACGGCTGGCGAAAAACAAATCGAGTCGGCGAAAATGTTTTACAATTAAAATTACGCG CGCAGCGGGCACAAAGGTACCAATATACACGAGTGCGAGCGCGAGTACGAGTATCTGCAAGATACACACACGTAGATCAA ATAAAGCAGCGCAATATGCGCAAATTAAACTAAAATGCAGCACATCAGAGCTGTAAAAACTTTTACGACAGCGACTACGA CTGGTAACTTGCCACCCGCACAGTCGAAAATAATACCAGATACAAATTAAGGTCGCCCAGCGGCAGCGCATAAACACGGT GGAACGGGAAGTGGCTGTGGAAGTGGAAGCGGAGGATTCTCTCGCCTCCTACCTGGAGTATATTAGTCTCAGTCCTTAGA TCCTTACACAAAAGGATAGCCAGCCAGGGAGCGCAGAACGCACACACAAGGGCCGTAAAACATTTTGGGCTGCTGCTATA CTGCAGGTCTCCTCCTCTCAGGACTCGTAAATTTCCAGCCCAACCACCACCACCGCCGCCCGAAAAACGGAAAATAGAAA CACCGAAAAATCCAAGACACCGCCAAGTAACTGCATCCTCAACCCGCTTATATATAGAACTATATAGGTACATATATCCG GGCAGCGATGGCGGGTAACATTGTCAAATGGTGGAAGCATAAAATTCTCGGCGGCTACAAACAATTCTCAGTCCAGGAAT GCACCACAGACTCCGAGGAGCTGATGTACCACCAGGTGCGGGCCTCCTCCTCCTGCAGCGCCCCGCCCGATTTGCTGCTG GTCAGCGAACGTGACAATAATATCCAACTGCGATCGCCGGTAGTGAACATAATCACCACGCCGCCGGGCAATGCGTCTGG TGCGGGAAGTAAGCAGCAGTCTCATCACCAGACGAACCACCACTCCTCGGGCAGGAGTCATCCCGGGCACACGGCACATC CGCAGGATGTGAGCAGCGGCGGCAGCCATAGCAAGCATCTGCGCATCAGCAGCACTTCCAATGGCAAGCACGGCAAATAC TCAAATATGCAGCAGCAACTGCCGCAGGATGAGGATGTGGTGGATGCGGCTGCCACGATGCAGCAGCAGCAGCACACTGG CCACGCCCACTCGCGCCACCTGCACCACCACAAGGAGGAGCGCATCCGACTGGAGGAATTCACCTGCGACGTGTCCGTGG AGGGCGGCAAGTCATCGCAGCCGCTGCAGTTCTCGTTCACGTTCTACGACCTGGACGGGCATCACGGCAAGATAACAAAG GACGACATCGTGGGCATTGTGTACACCATATACGAGTCCATTGGCAAGTCGGTGGTGGTGCCCCACTGCGGCAGCAAGAC AATCAACGTGCGCCTCACCGTCAGTCCCGAGGGCAAATCGAAATCGCAGCCGGTGGTGCCCGTTCCGGTGGCAGCCGGAT TCAGCAGCAGCCACGCCAGCAAACTGAAGAAGTTGCCCACGGGTCTGGCGGCCATGTCGAAACCCCTGGCCGGCGGAGGA GTGGGATCCGGCGGAGCGTCGGCGCTAACGACATCCGCCGGCAACCGCCGCCAGCATCGCTATCGACCACGCAAACTGAT TAAGTCCGATGACGAGGACGATGACAGCAACAGCGAAAAGGAGAAGGACGCCGCCCACGCCCCTGCCGCCGACCAGCCCA GCGGAAGTGGAACAAAGGCGACTGGGAAGAGCCATCACCACCAGTCGCAGTCCGCCAGGTATCACCAGAAGAACAATTCC CGGGCGGAGCAGTGCTGCACGGAACAGAATACGCCCGACAATGGCCACAATACCTACGAGAATATGCTGAATCTCAAGTG CTGCAAGCCGGAGGTGGACCAGGTGGACTGTCCCTCGCACCGACAGCACCACCAGAGCCACCCGAACCATCAAATGCGCC AGCAGGACATCTACATGAAGCAGGCCACCCAGCGGGTCAAGATGTTGCGAAGGGCGCGCAAACAAAAGTACCAGGACCAC TGCCTCGAAACGCGACAGCGCAGCCTGTCAGTGGGCAACGATTCCGCCTGCCCGAATCGCCATCTGCAGCTGCAGCAGCC GCCGGTGGGTCACCCCCAGCCCCAGTCGCTGAACCACAAGAGCGCGTCGGGGTCACCACCGCTGGGCGTGGGGGGTGGTG GCGACATGATGCTCGATGGGGTGCAGCTGCGTCAGCCGCGACCCCATTCCCTCACCCCGCATCAGCATCAACAGCAAAAT CAGCAGCAGCAGCAGCAGCAGCGAAAATCGGCCGAGTGCTGGAAATCGGCGCTGAATCGCAACGATTTAATTAGCATCAT CAGGGAGAGCATGGAGAAGAACCGCCTGTGTTTTCAGCTGAATGGAAAACCCCAAGCCAATGTGAGTCCCATACGGCAAC CGGCAGCACAACAACAACCACAACAACAGCAACGCCAACGCTGCAATACGGGCTCGAAAATACCCACGTTAATTACCAAC CACAGTCCGGTCGCCCAGCAGTCGCCGCTCAGCTGCAGTCCACCCACGGCGGAGCCCACCACCCCCAGCATTCCAGCAGC TCCGCCGGCCATCGAGGTCAACGGTCAGCAGCACCACCCCACTCATCCCACTCATCCCAGCCACCACAACCACCACGAGC ATCCCCAACCGCACATACCTATCTACCATCAGCAGTTGGCCATTAATCCGGCCGTTCTGGCCGCCCAGCAGACGCACAAC ACGGCCCACAACAAGCTGAATCTGTGTGGCTACGACTCCTTTCTGCACGCCACTATCTGTGGGGGCGGTGCAGCCGCCCA CTCGCCCCCGGCCACGCCCAGTAATGTGGCGACCGTTCAGCCGATACCCAAGAAGAGCCAGAAGAACCTGCTGCAAGGAT ACCAGCGTTTGGAGCAGTCGCAGCAGCAGCAGCAACAGCAGCGGAGCAGCAAGGACTACAAGAACTATGGCAACCTCATC TATGCCAAGCTGAGTGAGCAGCTGCAGCAGAAGGATCGGGAGCAGAGGCGACAGCGGCACAAGCAGCAGCAACACCAGAT GCTGCAGGATCAGCCCAAGGATGCGAGTCGGTCGGAGCAGCGACCACCGACATCAAACTCCAGTTCGGCTGGCTCCAAGA TCTACGGCGATGCCGTCGAGTGCGCCCATCTACTGGCCAGCGAGGAGGAGGACCTACCCCCCAGTCCGCAGCTGACCAGT ACGCCCAGCAAAGTGGTCAGCACGGACACCCTCATCGATCTCAACGACGATGTGGGCGAGGCTGTGGCCGAGGCAGTCAC AGAAGGAGGCAAGCAGTCGTTGGAGGCTGAGGAATCTGGCCAGCAGGTGGAGGTGGAACTGGACACCAGCGCCTCCAGCT CCATGATACACCGCTATGTGCACGAGCACATCCACCACCACTATCACCACTTCAAGGAGCAGCAGGATGTCTAGGCTAAT CGGGGAATCACACTATAAACTATTTGAATGTTGCTTATGTTTCAGTCTATTGATTAATGTAGCGCGAATTGTAATTTAAA GTAAGCCTTCATCAACTAAAACGAGAAAATATTCGAAAAACGATATTAGGGCCAGAAACAGTGGGGTTGGTTAAGTGAGT TTGGAGTTTGGTTAACCAAAGACAAATTGAGAACGATTGCTGACAAAAATCAATTGATAGAAATGCTAGATACTAAATAA AATAACTGATTATTATTCTTTGATTTTGTAAAACCAATATGTAAATTAGGATAAATATTAAACCTAACACAGTGAAGTTA AGCTAGGTTCCTGAATAGGTAAAGAGGTTAACCAATTTCAAACTCGCATCTCCAATTTCTCTCAAGGAAAATCTTTAGAA CCAAATTCGAACTCAATGAAGATTTAGCCCACTGTGAGATCCACTGATCCCAGCATATACATACATACATACATACACAT AGCCTCTATAGCTTACGCAGAATTCGAAAGAAAACAAAAATCATGCGCCTTTTGCCTTAGCAACAATCGAATCGCCGCTT CGCTATGCTACATGTTGCAGAAACACATATCACTATCAGAAACAACATGTTGCCAAGAGCATTTTGTGTTGCTGAGCGTG TACGTGTAAACTAATGGGAGAAAAACTATTGTTTAGTGCAAATTATTAAAACAAATTGTTGTCTATGATATAAAGTTTTA TGCGCCGCTTTTTACGTGTTTAGACAGAGAGCAACAGATGATTGTAGTGGAACTATGGCCGCCAGTTTAAGTTAACTACC GATATGGATCATGTATATTTATGTTATCTAAGCCAATTAATTTAAATACTATTCTTAGTTTCTAACACTAACCCCAAACA AGACAAGAAACAATGCAACTAAGTACTAAACCACAACGACCACCAACACAAATCGTAGTTCAATTATTATTTTAAAACTT GTTTTTAATTACATCATCTGTCAAACAATCGAGTTACCAATCCAATCAATCAAACAAACAAAAGAAGCACAAAAACAAAA TGCATTTAACTCTTATTTACATAAAAAAGACTGATTTCCAATTAAACAATATATTAATTAATTGCATATGAATAAAGGTA TATATAAATATATATAGACAATTTATGAGAAATACTAAATAAATTATTGAATTGCTAAAAAAAAAAAAAAAAAAAAA
Cloned cDNAs prepared from polyadenylated RNA isolated from larvae and early pupae.
Larvae and early pupae at various stages of development were collected from an isogenic y; cn bw sp (iso-1) strain.
RNA was poly(A)+ selected once. Oligo(dT) primed with XhoI site at end of primer for first strand synthesis; EcoRI adapter on 5' ends of clones; size fractionated on Sephacryl S-500--approximately 1-6kb. cDNA directionally cloned into EcoRI/XhoI-digested pOT2 plasmid. ~500,000 colonies were plated out and a mass plasmid DNA prep was done using Qiagen Plasmids containing the cDNA inserts were size selected on Sephacryl S-500 column in order to exclude plasmids with no insert. Early elutions were collected and transformed into DH5 alpha strain (Gibco/BRL).