PCR test for HT115/HT115(DE3)?
nobody at hgmp.mrc.ac.uk
Mon Jan 14 04:09:19 EST 2002
About using HT115 as a negative control, I am using it on a normal feeding
plates. It is still didn't prove a possible value comparing with using plate=
with OP50 as a control. However, I found that it might avoid differences
which could appear as a result of the feeding plate components.
Abd ElNasser Elashry
Lehrstuhl F=FCr Entwicklungsbiologie
III. Zoologisches Institut-Entwicklungsbiologie
Tel. 0049551395669, 00495519966779
Colin Dolphin wrote:
> 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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