http://www.alnylam.com/rnai_primer/rna-interference-pg5.htm. Effects of “knocking out” AGT-1 gene in C.Elegans by RNAi mechanism. Monica Coulson, Department of Biology, York College. Methods. Results. http:// wormatlas.org. Introduction
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Effects of “knocking out” AGT-1 gene in
C.Elegansby RNAi mechanism
Monica Coulson, Department of Biology, York College
bp Transformed Colonies
ladder 1 2 3 4
Figure 2. PR244 vector with AGT-1 insert. AGT-1 inserted into vector by BP clonase reaction. HT115 bacteria were transformed with PR244 vector and grown under Kanomycin selection.
Figure 3. Verification of AGT-1 siRNA fragment into PR244 vector. HT115 cells were transformed with PR244-AGT-1 plasmid. Colonies were picked and DNA was purified with AGT-1 primers as described in Figure 1.
Table 1. Phenotypes of wild type (N2) and siRNA AGT-1 C.elegans
100 bpC.elegans DNA
ladder RXN 1 RXN 2
Hau et al. 2007
Figure 1. PCR of N2 C.elegans DNA with AGT-1 primers. N2 DNA was amplified using AGT-1 primers (94 30”, 6030”,7230”) 30 cycles. Samples resolved on 2% agarose gel at 250 V for 12 minutes and imaged.
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I would like to thank Dr. Kaltreider and the Biology faculty for their assistance.