Selected article for: "dna extraction and restriction enzyme"
Author: Yang, H-C; Chen, T-L; Wu, Y-H; Cheng, K-P; Lin, Y-H; Cheng, M-L; Ho, H-Y; Lo, S J; Chiu, D T-Y
Title: Glucose 6-phosphate dehydrogenase deficiency enhances germ cell apoptosis and causes defective embryogenesis in Caenorhabditis elegans
  • Document date: 2013_5_2
    • ID: j3ku7i2c_17
    Snippet: Construction of G6PD-RNAi vector: L4440-G6PD. RNA extraction, competent cell preparation, plasmid DNA isolation, ligation, and transformation in this study were performed according to standard molecular cloning protocol. 48 In brief, the total RNA of wild type C. elegans was isolated using Trizol (Invitrogen Carlsbad, CA, USA) and reverse transcribed to cDNA using reverse transcriptase-Superscript III (Invitrogen). The full length C. elegans G6PD.....
    Document: Construction of G6PD-RNAi vector: L4440-G6PD. RNA extraction, competent cell preparation, plasmid DNA isolation, ligation, and transformation in this study were performed according to standard molecular cloning protocol. 48 In brief, the total RNA of wild type C. elegans was isolated using Trizol (Invitrogen Carlsbad, CA, USA) and reverse transcribed to cDNA using reverse transcriptase-Superscript III (Invitrogen). The full length C. elegans G6PD (B0035.5) cDNA was amplified by PCR with primer pairs (forward primer: 5 0 -ATGGCATGCAAACGTCATTC-3 0 ; reverse primer: 5 0 -CCCAACGAGGTTTCGA-TATT-3 0 ). The PCR product of full length C. elegans G6PD was purified and ligated to pCR2.1-TOPO (Invitrogen). The construct was validated by restriction enzyme digestion and sequencing. The full length C. elegans G6PD fragment was ligated to L4440 (L4440-G6PD), and later transformed into E. coli HT115 (DE3).

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