PCR test for HT115/HT115(DE3)?

Nobody 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=
s
with OP50 as a control. However, I found that it might avoid differences
which could appear as a result of the feeding plate components.
Nasser

Abd ElNasser Elashry
Lehrstuhl F=FCr Entwicklungsbiologie
III. Zoologisches Institut-Entwicklungsbiologie
Universit=E4t G=F6ttingen
Humboldtallee 34A,
37073 G=F6ttingen
Tel. 0049551395669, 00495519966779
Germany

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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