Comment: early pupae
GAACGAGAGCGGGAAAGAGAACACCTGTTGCTGGAGCAACAGCAGCAGTTGCAACTGCAGGAAGTGCAGGTGGCCAGGGA CAACCACCTCAGCCATTTGGCGGGTCAGCAGCAGCAGCAGCAACAGCGGAAGCGACACGTTTACGGCGAGGAGCGGGAAA GGGAGAGGGATCGAGAGCGTGACATGGATCGGGATCATCGCCTGGTCAATGGAAATGCAGATCCCAACGAAAGCTGGCAG CATCTGCGCGTTACGACCGAAACTTTGGCGGAGGAGGGCGGCAAACACGGCAAGCCGGCGGACAAGGCCCAGCACCAGAT GCACAATACGAACCAAAGTCAGAATTCGGATGTCAATGTCTACAGGGAGCACCAGCTGGACGCCTGGAAGCTGGAGCGCA GCTACACGCTGGCCGTGGAGTCGCGTCACGGCATCATTGAGTACGAGGGCTCGCCCAGACGCATTGGTGTAACTACCACC AGCAATTTGGGGGCAACTGGAGCGGCGGCGGCATCAGCAGCAGCGGACGAAAGCGAACCACCAGCAGCGGCAGAAGGTGC AGCTGCAGCAACAGCAACATCGCAACCACAACAGCAGGTGCCCATGGCCACTGTACCACCGCTGAGCTCCACGGCTACCT CGCTCCAGAAGACGGCCGCCCGCGCAGCATTGGCTGCTCTGGCCCAGAGCCATCCACACAGCTCCCACAATATACTCAGC ACCCTGCTTCCGCACTCTCCTAATCCAGTGAATCCCGGCCAGATGCACGCGGCGCAGCAGTCCATCCAGAACTATCCCGT GCGTCCTGGATTTCCGCAGCGCATTATCTCGCCGCCCAGTCGTGACCCGAGGAGCAGCTCCCCTCCTCTGCAGCAGCAGC AATATCAGCAGCAACAACAGCAACAGCTGCAACATCAGCAGCAGCACTTTGCCAGTCTGATGAGCAGTCACAGTAACAAC AACTCCAATACCAACAAACAGCCGCTGAGCAGCGACAATAATGCGGAGGACACCAGCAACGATGTGTCCGAGATTGGCAC CATATCCGACCTGACCACTCCGGAGGCAGTGGGTCTGTCTATTGGAATCGCTGCAGGCGAGATGGCCAGCCGGGCGAGCA CTCCACCGCAAACGGCACAAGCACCACCGACAGCAGTGGCGGGCAGCACTGCCAACGGAACCGGAAGCGGAAACGGAACA GCAACCGGAAATGCAACAGCAGCGGGCAGCGGATTGGGTCCCTTGAGCCTGCACACGAAATCATCGACCTTCGATTATCT CTACGAGTTCTCGGAGACGCGCAAGGTGCTGGAGGAGTTTTTCAAATGCCCCTCCACCGACGAGCAGCCCATCATGGAAA ATGGTAGCGATGTGGACAGCATTGACATCCAGTACGAGTTCCACAGCAACTTTGAGCGCGACGAGGAGGATTTGGCCGAG GAGGAAGAAGCCGAAGACGACGAGGAGGTCGACGACGATGCCGACGAGGAAAACGATGAGGCCGAGGATGAGGACGAGCC GTTGGAACAGGATCGTGACCAGAACTACCGCGCACAGGCGCCTCCCCAATCCGGCCACATCGATTCCGAGCGCCTTGTTT ACAGTGGTTATGGACAGCAGCCGCAATTGCAGGCACAGCTCACCGCTCAGCCGATGAGCGGGGCGAGACGACACTATAGC GGCAGTCCGCTAATGCGGGACTTTGACTTCTTCCTGGACTCGACCAGTCGGAGCAGCGGCGAGCGGGATGCCGACCAGGA TGGGCTAAATCACAACCAGCTGCTGAGCACGAGCAGCGGACATGGCGGAGGCGTGGGCGTGAGCGGTGGATTAGGCGTGG GACCGGGTGGAGGTCACAATTACCGCTATTCGCCGGAAACCACCGACTACGATTCCAACTGCGGCGATCTGGACAGTCTC TCAGGGGAGATGAATGGCGGGGTGTCGACCTCCTGCTCAAACTACGCAAAGTTCTATGCCTCGGCCATGCCGGTCCTGGA GGATGGCCTCTCCTCCGGTCACACCAGTGACACCGAGAACAACAACAACAACCAGAAGAACCTGCAGCAGTCGACGGTCA ACCAGGGTGGCCAGTGCCCAACCAGCTTAAGCGGTAGCACTAGCATTGGTGGCAGTGGAGGCATCTCCATGCTGATGGAC ATGAAGCGCATCTCGACGAACCACAGCCTCAACAGCATGAATAACCAGACCACCACCGCCTCCACGACAACGGATAGCAA TGGCGGCGTCGGCGGAGTGGGTCACCACCTGGTGGCCACACTGGTGGCCAGTCCCAACAATGAGCAGGCGAAGTCGCAGT GTCAAACGCAGCTGCAATCCCATCCGCCAAAGACATTGTCACTAGATCAGAGTCCCAGCAACCACAGTAAAGTGTTCAAG AACATTGATCCAGAGTTGGATTCGCTCTACTCGATCGGAGTCTTCCACCGTCCAGACACGGTGCAGATGACGCCACCTCC GCCCGCTCCGGCGCCACATCGAAAGCCCAACAATAATGGCTGCTCCAGTGGCGTTGTGAACACTAATGGCAGTCTAGCCG GTGGCAGCAACGACGTGGATTTGCTGCAGCCAAGCAGCAAGCACACTCCATTGGCGGCGGCCATTAGTTCCACGCTGCAG CGCAGTTCGCCCACTTCGACGATAACCGGCAATGGAATTGGAAACGGAAGTGGAAAGACCAGAAGCGCCGGCAGCAATTG CAGCAGCAGCGCCAACGGCGGCATGCCACCGCCCATTCCGGAGCGTACCAACCTCGTATCCGCAGTGCAAAGATCTCCAT CGCCTGGCCCCAACTCACCCGTATGGCTATCGCGCCATTTGGACGGCGGTGCTGGAAAGGATCTGCTGGAATCCGGCTCG GATAACCACTCAAAGAACCACAGTGCCGACGAGGAGGACGTGGACACCGATCTGGAGACGGACCGCCTGCTAGGCCACCA GAGGCTGGACGACCAGGGCTACTACGATGAGAACAAGAGCTGGGATCGTAAACCGCGCTCGCTGCTCTCGAAGATCTCGC CCAAGCAGCAGATACCGAGCACTAAGACGCGCAACGGCTACAATGCCCTGCTCAGCTCCACGCCGGAGCTACCGCCCCCG ATTCCGCCAAAGGGTCAGTCCGGATTGGGATTGGGTCTCACTCTGGGTGCCGGTAATGGTGGCAAGCTGCTCGACTTGGT TGGCGGCATGGTCCAGCGCAGTTTATCGCGGAGTTCGTCGGAGCACAGTGAGAAATCGCTCAGTAAAATGCCAATTTCTG GCGGGGATCTGTGTGCCGGCGGCGTGGATGTGGTAGCCTCAACTGTGGCGGCCACCTCGGCAACGGTGGTGGCCAGTACG CCGGCGTTGGGAAGTCCAGGCAACGGGGGTGGCTCGGAGAGATCAGCTGCAGAAATGGCCAATCCGGGTGGAGCCGTTAA GCTGAGCGAGCAGGAGATTGGAACCATTAACGGTGGCAGTGTTATGCCATTGGGATCAGGATCGGGCACTGGATCTGGAG CAGGATCGGGAGCCGGATCTGGAACGGGAACCAATGCAGCTGTGGTGGCCAATAGCAACGCCAACAACAACGTCGGCGGC GGCGGCGGCGGCGGAAACAACAGCAGCGGCAGTGGCGCCGGCAGCAACAGCAACAGCGGCGAGAAAAAAGTGAAAAAGAG TAAGAGCAAAGAAGGCGGTGCAGTGCTCATCGAGGGCGTTCTCTTCAGGGCCAAGTACTTGGGCTCCACGCAGCTGGTCT GCGAAGGTCAGCCCACGAAATCAACGCGAATGATGCAGGCGGAGGAAGCCGTCTCCAGGATAAAGGCGCTGGCACCCGAT GGCGATGTGCAGCCCAGCACAGAGGTGGATCTGTTTATATCGACGGAGAAGATCATGGTTCTGAACACGGACCTCAAGGA GATCATGATGGATCACGCCCTGCGCACCATCTCCTATATCGCGGACATCGGGGATTTGGTGGTTCTGATGGCTCGTCGCC GCTTCGTTCCCCAGGACATCGATGATGCACCCAAACCGAATCGCACGCCCAAGATGATCTGTCACGTGTTCGAAAGCGAT GAGGCGCAATTTATTGCCCAATCGATTGGTCAGGCCTTTCAGGTGGCCTACATGGAGTTCCTCAAGGCGAATGGCATTGA GGATCATCGGTTTGTCAAGGAGATGGACTACCAGGAGGTGCTCAACAGTCAGGAGATCTTCGGCGACGAACTGGAGATCT TTGCCAAGAAAGAGTTGCAAAAGGAGGTGGTCGTTCCAAAGGCCAAGGGCGAGATTCTGGGCGTGGTGATCGTGGAAAGC GGCTGGGGCTCAATGCTGCCCACCGTGGTAATTGCAAATCTGATGAGCTCGGGAGCTGCTGCCCGATGTGGACAGCTCAA CATCGGCGACCAGTTGATCGCCATCAATGGACTGAGCCTCGTGGGTCTGCCGCTCTCCACCTGCCAGACGTACATCAAGA ACACCAAGAACCAGACGGTTGTCAAGTTCACGGTGGTGCCATGTGCCCCAGTCGTCGAGGTGAAGATCAAGCGGCCGGAG ACCAAGTACCAGCTGGGATTCAGCGTCCAAAACGGAGTGATCTGCAGTCTTTTGCGAGGTGGCATTGCGGAAAGGGGTGG GGTTCGTGTGGGTCATCGCATCATCGAGATAAACAACCAGAGCGTGGTGGCTGTGCCCCACGAGAAGATCGTAAACTTGC TGGCCACTTCAGTGGGAGAGATACTCATGAAAACGATGCCCACGTCCATGTTCCGCCTGCTCACTGGCCAGGAAAATCCC ATATATATTTAAGCCTGCGGGAGAGTAATAAATGGCGATTATTGTTAACTATAAAAAAAACACAACCTTACAAGGGTATT ACGATATTTAGTATATATACCTATACATATAGAGAAAGAAAGTGGAAGAAAGAGTAAATATATGTATGTATATGCGTGTG TACCTATATATATATGCTAAACATATACTACACACAGGTGTATCGATTTTTTTTGTGAGTAATACTAGACAAAGTTCGAT CGGAGGCAAGTTAAGATCAGCTACTACGACAAATAAGAGGAAGCGATAACTAACACACACACAAACACATAGCACGCCTT TTTGCAAAGCTTAGTAATAATAGTTAATAATGTGTGTGAGTTTGGGCCATCGGCCAGGACATACATATATACATACATAA ATATATATGAACAGAATGACCTTTTTGCGAAAACGTCCTTAGCTTAGTTGAACCGCGACATTAGTCAGAGAGTACAGCAG CTATAGTAACAGAACCCCCATATATGCTTACCCATACTTATTGTAAAAAACCAAAAAAAAAAAAAAAGAAATCCACAAAA ACCATGGCAACTCAAACAAACCCAAACAAGATTTGAATTCAGATACGAGACAGGCAGCTCTTTTGATATTGCATATATAA TCCACACACTATATAGCACCATCCCGAATATATTTATATAAACAAACAAAAAAGACAGAGGGGGGGGATGTGGAAAAGCC TAGGACAGATGGGAAATGGTTAGAGAAATGGAAATGAAAATGGATATTGTAACTGCAGCGTTTCGTTTTCTACTCGCACC CATAGCCGAGAATTTTGATAGTTATTAAACGGATATAGCCAGATCGTAATGTATGCCATGTGTAAATTAAGGAGCGCTTT ACACCCCCACAAATACGAACATATACCCCCCATAGAGGATTTAGTTTAGCAAACACTTACACTCACACTCACTTAGACGC TTCGATTTCAACCATCTTCGTCCAAAAATTGTGCTACTACTCGTTAAACAAGTTGATAGATATAAAAGGGAATAAAAAAA AGGATAACAGCAGGAGTAGGGAGATTGACATAGAGACCCCCGTTATAAGCCATAAGAAATAGAAAGAGACGGATTAGCTT AAAATAACGTATATCTGGGTTCGCATTTTAGAGCTACTCGAACGGAAAGAGACGGCAACAGGAACAGAGATAGAGACAGC TATAGAGACAAGCAGACAGCTAGACAGAATGGGAAAATTGCATTTCTTTGTGTGTATATGTATGTGTAAAATACTCGCTA TTTGGCCCCCCAATCGCAATCCAAGCAACCGGCAGCAGCAGTCACTAAAAGAACTATTATTAACTATGATTTAATTATAT AACGGCACCGGGGCATTTGCTTGATGTTTGTGAATTGAATGACTATTTGGCCTCTTTTCCGCTGCTTGGCAAGGGCCACG CCCACGCCCAAAAAAAAAACATAAACAAAATTATTATACAAAAAAAAAAAAAAAAAA
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).
