Comment: early pupae
CGGAATCAAGTTTATCGCAAGGACGAAGACGCTCCACGTTTTTTTCCCGGTCCGGCGCGATGTGAGCAAGTCCTAGAGAA CGCGCGCGAAAACTTCCCACCCAGTCCACCCAACTCCACCCCCCCCCCCCCCCGAAAATAAAATCATCGGTAGCTTTTGT ACGCCGTACAGGTGGAAAGGCGTAGACAAGGAAACGGGCAGTGAAATCTATAAATTGACGCAAAAGTGAAGTGAAGTGAA GTAATTTAACCACGCCCTAAGCACCAAAAATCAAACAAACCAGACATGGAGCACTTGCGACCGCTGAGCTACGAGGAGAT GTGCCAAATGGAGCTGCTCCAGGCAGCCGGCGGATCAACGACATCCTCGGCTCCGCCGGCCATGATGCCTATCATGGTGA TACCCACAGCACTGCTGCCGGCTCCCATCGACGGGGCAGCGGGCGGTGGAGTTCCCGGTGGTGGTGGCATTGGTGGTGCG GCCATGGTGGAGCACTATTATCAGCTGCAGCCCACGCACCACCTCCTGCCAGCCGAGCCGTTACCAACGACGGCGACAGC AACAGCAACAGCAAATTCCGGCGAATTGGCGCTCTTGCAAGGCGAACAACAGATTCCACATCCGAGCATACAGCATCCAC AGCAAATGAATATCCTGTGCACCGGAACACTGGGACGCCAAAGGATGCCGGCATCGGGTGGTGGCGGGCATAATCCAAAC ATAAATCCCAATCCAAATCCCTCGGTGGCGGCTACTTTGCCGCGCGGTTTCCAGCCCGCCGCCTTCGTTACGGACGTCGA GTCGGATTTCGAGGGCAGTGAATGCGGTGCGGGCAGCATGACCACCACCTGCATTCCGCACTTTCGCTTCGGAGCGTGCG GCCTTCAGTCCCTGCAACAGCACCACCAGCAGCAGGATGCAGTGGTGGGCTGTGGTTCGCTGGCCTACATGATGGGCTCT GGCACCTTGGGTCGGGGCCGTCGTCTGGCCGGCGGTCAGGGCAAACCTAAGCGGGTTTCCTTCAAGGGCGACAACCTGCC ACCGCCGTCGCCACCGCCGCCACCAGCCGTTGAGCTAGACGAGAATGGGCTGCCCATACCGGGTGGACCGGGAGATGCGG GTAGCTGCTCCTACATGCACTTCGACAACTACATGGACTACCAGACAACTGGTTTCGGTGGATTGCCCGACGTCACCGAG CACTCGAACATCCGCCGCACACTGTCCCGCCAGAACTCGATCTCGAACAGCGATTCCGGCGGCGAGATAGCCAGCATCCA GAAGTCAAAGGTGAACTTTGGCAATCCCCTCGCCGGCGGCGTGGTCGTCGGTGGAGATATAGTTGGTCTGGACGCCAAGT CGCCCACATCGCTGGGTTCCACGACTGGAGCTGGCGCCACTGGTAAAGCGAAGGGAAAGGCGTCAAGGAAGGCGAATGCC GGACCAGATGGCGGACAGAAGGGCAGGAGAGGATCCTGGCTGGTGGCCATCACCCTGGTCAGCGCCATCTGTCTACTGGC CATCGCCGCCACTTTGGCCTACCAGCACTTCCTTATGGCGCCCAGCCGCAATTCCCATGCCCAAAGACTGAGAATCGTTC GTCGAATCCTGCGTGAGGTGCCGCTGGTCGACGGGCACAACAATTATGCCTGGAATGTTCGCAAGTACGCCCACAGCAGC CTCGAACTCCATCTCAGCCACGACGTGGACCACAAATCACTGTGGTCGCGACCGGCGTGGGCGCAAACGGACATGGAGCG ACTCAAGCAGGGATTAGTCAGCGTCCAAGTGTGGTCAGCATATGTGCCGTGCGAGGCGCAAGGCTTGGACGCGGTCCAGT TGGCTCTGGAGCAGATCGACATTGTGCGGCGATTGTCGGACATGTACGCCCGGGAAACGGTGCTGGCCACATCCTCGCAG GACATTGTCGATGCACATCGCCGGGGACTCCTGGCCTCGCTAATTGGCGTGGAAGGTGGCCACACGATTGGCTCCTCCCT GGGAGTCCTGCGATCCTTTTACTCACTGGGCGCACGCTATCTCAGCCTGACCCATCGCTGTGACGTCTCATGGGCAGGAT CGAGTGCATCGCCGGCGGAACAGGGACTCACGCCCTTTGGCAAGGCGATTGTGCGTGAGATGAACCGGCTGGGCATGATG ATAGATCTGTCGCACAGCTCGGATGCCACGGCCAGGGATGTACTTCAGGTGACCCGCGCTCCGGTCATCTTCTCTCATTC CGCCGCCCGCCAGCTGTGCAATTCGACGAGAAATGTGCCGGATGACATCTTACGCCTGGTGGCCGAGAACGGGGGTCTTA TCATGCTGAGCTTTGACTCCGAGGATGTGGCATGTGGGCGGCAGGCGCGCCTCCAGGACGTGATCGAGCACATTAAGTAT GTGCGGGCCATTGCCGGCATACAGCACATTGGATTGGGAGCCGGATACGATGGCATCGAGCTGCCGCCCTTGGGACTGGA GGATGTGTCCAAATATCCCGAGCTCTTGGCCGCCCTGCTGGAGGATCACAACTGGAGCGAGGAGGATGTGGCCATGCTCG CGGGCAGGAATTTTCTGCGCATCATGGAAACGGTGGAAACGGTGCGGGATTACTGGAAGCGAGCGGCCATCCAGCCCATT GAGCAGACGGAGCCGCAGCCCAAGACGCAGTGCACCTACATGTCCTCGTGACCGCAGGCGCAGGCGGTGAGGCAGCGACG CGATGAGCCGCCCATGGAGCGGGCCAATCGAACCACTGCCACCTTGACGGGGGAGCACTTCTGGCAACGCGGTCCGGAGT CGAGGGACTCCGTCTCCAATCGCACCAATCCGGCGGTCATCGCGGCCACGACAACGAGTTTTACGGTCAGCGAACTGTTT CAGTAAACTGGAATTAACATAAGAGATTTTGGCTTGCGGTTCGGCTCGCCGAGATTTCGGGAAAGGAGGGCGTTTACTGA GGTACTGAAAACCCGGGAAGGGTCTTTAAAACAGTTCAATTTCAAGGGAATACTTTTTCTTTAGATATTCAATATTTCAT TATTTTCTTTGGGAATGAGAAACACATATTGATAAAGCCAATTGGAAATTCGTTAAAGAAGCCAATATGTACATAAACTA GATCCTTTGTGCTGCCATCACTTGCCCCCTCTGGTCGAAAAGTGAACGAAGAACTGTCGTGGCGAACCTCGTGAATAAAT AGTGTGTACTCGTAATTTACTTCCATAGGTTGCATTGATAGGCGTATGAATAAATAATAATAATAAATGTACTAGTATTT AGCCCATAATTGTTTGCAGCTATACTTATTTGGTGTTGTGGCAATGCATAATTTATGCACAATGCGGATACTTATTGATG TTGCTGTACCATGGGGCGTATGCGCGACGCAGCGAGCACCGAACCGCAAACCGAAATCGTTCTTGGCTTGCCTGTGACTG TATTATATATGTAGGGCACACATCAATGATTCACGCAAATTATCGCAATTAAGCGCAATATATTTAATCACAAACATCAC ACATACATAAATACAAACGAATACTTGTATTAATCATCGAAACGAATTAATTGCAGCAGTTGGCATTTATGCCAGGATCA TAGGGTCAAACAACATACAAAACTGTAAGCAACGCCAGCAATTCAATTAAAATGAGATTTTTCACCCTGGCTGCATGTGA ATCCATCTGAATATCTGTATTAGGTTAGTTAACAAAACACAAATGAAAAATCGGAAAAACAAAATCAAAAAAAAAAAAAA AAAA
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).
