This is a good thought! Thank you - trace amounts of the expression plasmid would not have crossed my mind. I am not sure whether I'd have enough DNA to cut out of a gel/clone/sequence (the bands are very weak), but I can certainly try...Magda
From: Cathal Garvey [mailto:[email protected]] Sent: Sunday, 19 September 2010 9:40 p.m. To: Dunowska, Magda Cc: [email protected]; Peter Ellis Subject: Re: RE: non-specific bands in NTC Hi Magda, It is possible that you are seeing amplification of the trace expression plasmid from production of the polymerase enzyme. To rule it out I guess you'd have to sequence a large band and blastn the result? --- Twitter: @onetruecathal Sent from my beloved Android phone. On 19 Sep 2010 02:57, "Dunowska, Magda" <[email protected]<mailto:[email protected]>> wrote: Just to clarify my previous email: NTC is "no template control" in the PCR run. It contains no template, so it is a type of a negative control. The difference between NTC tube and other negative tubes is that NTC has no template nucleic acids at all, while other negative samples have template nucleic acids in them, they are just negative for a specific sequence that I am looking for - I am looking for viral sequences in animal tissues. As I specified in my original email, the NTC tubes contain exactely the same master mix as all the other tubes - this is why I am so puzzled by what I see on the gel...Sorry again for not defining the abbreviation - Magda ________________________________________ From: [email protected]<mailto:[email protected]> [methods-boun...@... On 17/09/2010 15:09, Nick Theodorakis wrote: > On Sep 17, 7:59 am, > WS<[email protected]<mailto:[email protected]>>... _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
