Comment: early pupae
CAAAAGCGATCCCAGCTCCTTAAACTCATTCTCGTCAAGAACCACAGCCACATCCTAGCCCATCCTCCAGCGCAGCGGGA AGACGAGGAGCGCGGCGGTGGCGAGAAGGAAGCTAACGCTAATGAGGATCTATTGGCAGCATGAGTACACTGGGGGGAAG TAGGGGAGAGCGAAATGCCAAGCCCAAATTCACAGCGTTAGATATAAATCGCATGTACAAGAATAGCCGCGGCGAGTCAA GTGAACCATCTGCACAAAAGAATCAAGTGCCACGTAAACATGGAATGCAAATCTTGGGCAAAGTGCCGTCCGCTAGGCGA CCACCAGCCAATTTGCCATCCCTGAAGGCCGAGACCCACACCAGCAGCAACAACAATCTCTTAGTCTCAGTCGAAGGAGC CACATCTGGTGGGGCGGGCGCAGGCGGAGCAGCGGGATCTGGCGCGGCAGGAACCAACACTAGCCACAACAACACCACTG CCCACAGCGGAAGTTCAGGAACGAGCGGCGGAGCGGGATCAGGCGGCGGAGGAACTGCGTCTTCATCTGCTGGTGGCCAT CTATCCCAGCAGCTGACTCACAGCCAGCATCAGCACAATAATAACGTCGCTCTCGGCCCTAACGCCGGAAAGTTTGTCAA CAGCAACAACGGCAGCAGCAACAACAACGCCGGCAGCTCAGGCAGCGGCAACAAGAACTTCAAATTAATCAACAACTCCG TGGGCAGTTCCGGCGGAGGCGGACAAGGAGGGTCGGGCGGCGGCGGATCGTCAGGAGTAGGAGGTGGATCAGGGTCAGGA TCGGGCGGACACTCAGTCAACTCACCCAAGACATGGTCTGCAATTACCACAGGACACGAGCGGGGCTCGGGCGGTCAGAA CTATGGACGCCAGCAACAGCAGGTGGTCCCCCATTATCAGAGTCCACAGTTCCAGTACGAATTTCCCACGCTGGACGGCA CAGTTGGCAGCGGCGCCGGCGGAGGATCGGGAGCGGGTGTGGGTTCAGGATCTGGAGCGGGCAAGAGCCATTACCAGAAT CAGGTGCACCATGCCGGCCACCAAAACCATCACCAGCATCAGCAGCACCACCAGCAGCATCAACAGCACCACCACCAGCA GCAACAGCACCATCGGGACTACCACCATGGCCACGGGCGCCAGTATGGACACCACTCGCATCGCGACTACAGAGATGGCA GCGACGACGTCGGCGGCGGGTCCGCGGATCTGGGCGAGCTCAGCCTTCGGCCACAGAACGACACGGCCGCCTGGTTGCAG CAGCAGGAGAAGGCGGCCAAGGTGGCAGCAGCTGCGGCAGCGGACCAGGGCCTCAACCTACAGGGCCCGGGCCACCATCA GATGGGCGGTAATGGCGGAGCAGGAGGAGGGTCGGGCGGAGCTGTGCCCCTGCCCATACTATCGCTGATGCCCTCGTTCA TGCGTAGCGGAGCTCCGATGCCCGCCAGTGGAGCGACAGCGGCCAGCATCTTGGCCGGCGGATCGTTGGACACTTCTGCG TTGCCCAACGCTCCCTACCAGAACGGAATCCTGGGTGCTCGGTCTGCCTCGCCAGCTGTGGTGGGCAGTGGAAGCTTCAG GGCGGCCACTGCCATTCCCAAGCGACCACCGACCTCCTCGTCGCCCCCACAATCTGGCCTAGGCATTGGAGGATCACCCA CTCCCCCACAATCCCAGCAGCAGGCCAACGGAGGCGCTGGCGGAGCCGGGCGCAAGGATAAGGACTACGTGGTGGAGCCG GAGGTGGCCCAAATGCAGCGTCCTATCATACGCGAAGAGGACTTGGAGCGCCTCAACGCCATCGCCAAGGACGATAGCTG GACCAAGCAGGACGACATTGACTTCACGAAGAAACTAGCATTCTCGGACGATGAGTCCACTCCGGAAGAGCACCAAGGCC AGGGCAAGCAGCACAGCTCCGCCCAACTGCAGGCGCAAGGATCCTCCTCCTCAACCGATCAACGAAAGCTGTCCGCTTCC ACAACCAGTTGGCAGCGCGGCAAGGAGAGCACCGATCGCGACCAGCCATCGGAAGGCATTCAGGATCAGCAGCAGCAGCT TCAGCAGCAACAGCAGCAGCAAGAGGTTCGCCAACAGAATGGCCACCGCGGTAGCTCTGGCAACGTTGGAGCACCAGTGG CCGGTGGAATTGCCTTGGATGCCAGCGTGTACGAGCGAGTAAAGCAGCGCAAGGAGGAGGAAGAGCGTCGCGAGATGGAA CGCAAGCAGGCGGCTGCTAAGAAGTTGCAAGAACTGGAAATGAAGATGAATAGCAAAAAGGCAGCGGCTGCGGCTCTGGC TGGAGCCGAGGGCGGCGTGTCCTCTGCCTCAGTCCCGTTGAGCAATGAGGTGCCACCCACGTCCGCACCGCTAGGCAATG CCAGCGAAGATGAGGGCGGAGGCCAAGGCGGCGGCGGAGGGTTGAATCGTCGACCCCGGGGAAGCATCTCTAGCTTGGGA AGCGGCGTAACACCCACGGGAGGAGCTGGCGGCGGCGGCGGAGGAGTTTCGTCATCCTCGTCAGCAGGCGGCGACTATGG TCAAAAGAACTTCCTTACCCATTTCCAATCAAATTTACCACCACGTTTCCAGCGCCAGCAACAGCAACAACAGCAGCAGC TGCCGCGCTTGGAGAAGAGCGCCTCGGCAAGCAGTGCATTCGATAGCAATTCCCGCTACCTCCAGAAGGGAGGAGTAAGC GGAGGAGGCGGTGGTGTAAGTGGAGGAGCGCAAGCCAATGCTGGCCAGGGTCGCAGCATCTCCGGCGGTTATGTGCAGCG TGGTGCAAGTGTCGGAGGAAGCAGCGGATACGGACGAAATCGTCACGACAGCAGCAGTGCTCGGGAGGATCAGGATCTGG ATCAGCAGCGTTTCACCCGTGGCCAGGACTATCGGCAGGCACAAGCTCTGCCGCGCAGCATATCTGAGAACTCGCACCGT AAGACCAGCGTCTCATCGGCTGGCGACGATGTGCTGGGTGGCTCGTGCTCCTCATGGGCGGAGCAAACGGATGCCGAGCA GAAGGTGCACCATCGGCACCGCGAGGAGAGCTTCTCCTCGTCGCACAGCCATGACTCGGCAGTGCAAATAAAGATCCTGC AACGCCCGGCCCGACAGCCGAGCCTCAGTGAGGAGTCCTCGACCCAGCAGAAACAGCTGCCCGCCTCGCCGCAGCAGCAA AAGACGCTGACGACAGATCCCATGCCTACCCAGATTCTGCGACGCAGCGTGGATATCGATAAGTCCGGTGATGAGCGCGC GGAGGACAAGTCTGCATTGGAATCTGATGACAAGGCGAAGTCGGCTGGCAAGGTGGAGGAGGACAAGAGCGGCAATAAGC GGGTAAGTCCCAGGAGCGGAGCTGCTTGCGGAGGTCGAGGTGGCCGTGGTTATGCCGGCAGCTCTGGTGGATCATCCGTC GGATCGGGCAGTAGCTACCGCGGCCAGAGAAATGCCAGCGATTGGGGAAGCAGTCGTGGAAGCGGCGGCGCCGGCGGCAG GCGCTACTACGGCAGCGGCGGTGAGCAATCGGAGCACTCCGAGGATGTGGACGAGGAGTGTTACAGCAGTGGAGGAGGCG GAGCCGCTCGTCGCAGCCCAAAGGAATCGGGCAACGGTGCGCCCAATTCCGGTGCGTCGGCAAACAAGGCAGGATTCTCG CCACGCGGCGAACCATCTAGACGTGGTCGAGGCGGCCTCTCTAGCGGAGGAGGCGCCTCTGCTTCTTGTTACCGTCGACA GCCCGTGGGCACTGGATCCGCTGGTGGATCTGGTTCGGGCGGATCGGGCCGCAGCTATGGCAGACTTGGCTACGAGGAAT ATGGCAAGCAACAGCGAACTTCCGAGTCGGAGACGGACCTAGACAAGACCAAGCAGAACCAGCTGGCACTCAGCGCCGGC CTGCACAAACCAAGCAAGGATGACAAGGATTTGCCTGCGCTTGGGGGCGAGGAGAAGGACAAGACTGCCGCCCTGGCGCC TGCCCATAACACTGCCCTGCTAGCAAAGGATGAATTACAGAAAGATGCAGCGGGTCGCCTGCCGTTGCCTCCCGGCCTCG CAGCACCTGGCTCTGCAGTGGGATCCGTTACTGCCACATCCTCATCGGTCGTAGTTCCCGGTGCAGCGGACAAGAAGAAG ATTGAATCCTGCGCCGTTGTCGGCGGTGAGAAGGTGACGGGTAGCAAGTTGCCCGCCATTGGGGAAAAGACTTCGCTCGG CGCCGTTGGCTCGGGTCCATCAACCAGTCTTCTACTGGACGCCGGCGCACCCGTCAACACGATTATATTTGAGAACTCCA CGTACAAGCAGCAGGCAGCTGCCGCTTCCGTGGTAACCAAACCCGGAACGCTCTCGGTGTCAAGCAGTGGCGCCACCATG ACCGTGGACAGCCTGTCCAGCGCACTCTCCCAAATGAGTTTCGGCGGCAAGGCGACGGCGGCGGGCTCTGGCGATGATTC GCAGGATATGAAACTAGGATTTAGCTTCGGTGACGATCCCACCACACCCCTCAAGGTTGTGGATAACATCGATAAAACGG CCAGTCAGCAGCAGCAACAGCAGCAGCAACAACACTCTCAACAGCAACAACAACAGCAGCAGCAGCAAAACAATTCGACG GCGGATCTTAACATGAAGATAGCCAGCGTAAAGAAGGTTTGGGAGTCAGCCACGCCCATGTCCGACGGCCCATCGCCAGC ACAGGTGCAGCAGCAACAGCAACAGCAGCCCCAACAGCAACAACAGTCCCAGGTGCAGGTGGCGGCGCAGCAGCAACAAC AGCAGGTGGTGGGACCGCAGCAGGCAGGTGACGACGGCAGTGGTCACATGAGCGCCGCCTCCTTTGTGGCAGCCGTGGCC GCCTCACAGCATCAGCAGCAACACCAGCAAATGCCGCACTACGCGGTGTCGGCGGTGCACATGCAGAGCCACCACCACCA ACAGCAGCAGCAACAGCAGCAGGCAGCGGTGGCGGCAGCGCAGGCGCAGCACCACCAGCAGCAACAGGCACAGGCGCACC ACCAGCACCATGCACTTTCCTCGCCGGGCCCGGTGCCCGGTAGTCATGGCCATGGCTACAGTCATGGATCGCCGTTCGAC GCTGGATCGTTGGAGCAACAGTTCCAGCAAGAGGATTATCCCAGTCCACAGCAGCAACAGCAGGCGCACCAGAAGACCAA GCAGTTGCAGCAGCAGCAGCATCTGGGCATGTCACCGCCGCCCAGCCAGCAGCAACAGCAGCAGCACTCGCAACAGCAGC AGCAAGCACAGCACCAGCAACAACAGCAGCAGGCCCATCAGCAGCAGCAACAGCAGCAGCAGCCTTCATTCTACCAGGCG TCACCGCAGTTCGGCGTTGGAGGCATTCCCACTATCCCCAGTCCGCCGGCCGTGGTGTACAACTCGTCGCAGATGCCGCC ACCGTCGCAGGGGGGTAATCTGTACGCTCCATTCCATTCGCTCGATCATTCGAGCCGTTCGCCCGCCTACTCGGCGGCGG CGGCCGCCCACAGCTACCCGAACCCTTATGCGGCAGCAGCAGCGGCGGCGGCGGCAGCGGCCGCTGGCGGAGGTCCCTTC AATGTCAACGTGAACGCATATGCGATGCAGACTCCGCACGGCGGCAGCCTACCGCCCACAGCGCCCACACCGGACATGTA CTCGAACCTCTCTTCGCAGTTTCGATTGGGCGGTGGTCCGGGTCCCGGACCCGGTCCCTTTGGCCAGCCCAGCTCGCAGC AGTTAAGCAATCCGAGCAATCATGCTGCGGTGGCTATAATTTCGTCAAGCAGCAACTCCATGATGTCGTCAGGTGCGGCC AAACCGCCGCAGAGCCAGCAGACAATCGGTGCCATTGGCTCCAAGTCGGCAGGTAGCGGTGGCGGTCCAGGACCAGGACC TGGTCCCTATGCCACCACCCAGACGTATATGAATCTTTATGCGGCTCCACCGCCGCAACATCCAGGACAAGGTGGACCAC CGCCGGGTGCCCATCAGCTGCAGTCGAACAGTTTCTACTCGAATTCGGCGCCAGGCGGTCCGAGCGGACCACAGTTCTAC GGTGGTCCGCCGCCGCAGGGCAATGGTGGAGCGCAGAGCTATGGACTGGCCACCGCGGCCGGCATGTATGGCGGCCATCA GGGCGGACCGCCCGGCTCCAATGGGCCGCCGGGACCGCCGCAGTCGCAGCACACGATGGGCACGTTCAACACGCCGTTCA TGAACTCGCAGCTGCTGACGGCGGCCAGCATTAATCAGTACCGCGGCGGACCAACGCCTGGAGCGGCTTATCTCAAGGGT AACCAGGGCCAGACACACATGCAAGACTCGATGGGTCGTCAGCTGAAGAGTCCGCTGAGCGCTGACATGAGCCTCGGTCT GGCCAAGCAGGTGCAGTCGCAGCCCAGTCCACCGCATCACAAGAACTACGGCGCGTGGGATCTGCAGAACCAGGTGCTTC AGCAGCAGCAGCAGCAATCTCAGCAGCAACAGCAGCAGCAACAGCGCCAGGTCGGCGGCGGCAATGGTCCGAGCATGAAC GCCTTGGGCGGTCGCGGCAGTGGAGCGGTGGGCAGCGGCAGCGGCAACGGCGGAGGAGGAGGAGGAGGAGCTGGTGGGTC CCAAGTGGGCGGCAATGGCGGCGGTAACGGCGTTGGCAGCGTGGGCCAAAGTGGCGGCGGCGGCCAGGGACGCTATCCCA CGCCCATTCAGCGGCCCAACAACTATCCGCAGCATCCGCAACAACAGCAGCAACAACAGCAGCAACAGCAACAGCAGAGA GAGCAGGCGGCAGCTGCCGCCGTGGCGGCTCAGCGGGCACAGAATATGCGACAGGTGACCGGTGCCGGCGGAGGCGTGGC CGGAAGTGCAGGTGGCGGTGGTAGTCAAACACCAGTGGGACCGGCCGGCGGCAATGGTGGCCCCGGTGGCGCAGGAGGAG GCGGTGGAGCTGGCGGTGCAGCAGGATCAGGACCCGGAAAGCCCTACTATGCGAATAACGCTAGCGGAGCGGCCGTTGCG GCCAATCGAGGTGAGTGGCATGCGAACTGAGTGCCAGATGGTCACCACCCATCGACCAGCCGATCCCAGCCACCCACCAT AACAAACCCGGAATAACTATGTACGAGTACGAAACAGAAACACAATGATGCAAATTGATAACGAAGGAGGATGGCCAAGA GAATGAGCAACAGCTGGAGGAATGTATTGCATCCAAAACGAGAATATGAACGTAAACAGAGCAAGTCAGGAGTAAAACTA TATATGTGGAATATAATCAAGTTACCGAGCGAGACACAGGCATCCGAATAATAAGCCCACAAAATATGTGAAAGGGGCGA TGCGAGTAGAATAGTGCTATAGCTGTTTGGGGGAATCCCCGTCACAATCGCAAAATGGCAACTCAGCTAAATCACCGTAG CGAGGAGGAAATTAATTTAAATCGCAATTTGAAAGTAAGAAAAAAGTTAAGACGAAAGTTAAGAATCAACAGGAAACGAG TAAGGAATACCTAAATGCAAGAAGAAATATTTAACAAAAATGAACAGAATACAACACCATGCCACACCACAACACACACA CACACACACATGCATACATTTACATATACATACATACATACAGAATAGCAGATTGTATTTTTTGTCAAACAAATTTACAT AATAATATATTACTTTTATATACTTAAAAAAAAAAAAAAAAAA
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).
