PCR test for HT115/HT115(DE3)?
colin.dolphin at kcl.ac.uk
Fri Jan 11 08:11:22 EST 2002
Does anyone know the sequences of a primer pair that can be used in a
test PCR (using chromosomal DNA as template) to ensure that HT115(DE3)
RNAi feeding bacteria contain the DE3 lysogen. ie. are in fact HT115(DE3)
rather than HT115? These bacteria are often referred to simply as HT115
rather than HT115(DE3) which, considering the important differences
between them with respect to RNAi, could easily lead to problems and also
just seems rather sloppy. Also, do people actually use HT115 as a
negative control in feeding expts? If feeding RNAi was performed in both
HT115 and HT115(DE3) any observed phenotype in the latter is very likely
to be due to the expression of dsRNA, and hence an RNAi-mediated effect,
rather than other non-specific one(s)?
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