I apologise, I didn't mean to make it sound like a UK "consensus"! Just a consensus of the people still in the office at 5pm! I should clarify that our team works primarily with modern DNA and we do not have any trouble with our reactions omitting BSA. I believe the high-throughput sequencing teams do use BSA as part of the commercial kit; I am not sure why it is required in their chemistries but not our small-scale experiments. I suppose modern samples are more simply cleaned than aDNA so inhibitive compounds are not such an issue.
However I stick by my original statement, that if a paper details the contents of a PCR reaction and does not mention BSA, then BSA should not need to be assumed!
Susannah
Frederika Kaestle <[log in to unmask]> wrote:
> Actually, we still use BSA a lot in the US in our ancient DNA work. In
> the past (when I was in grad school in David Glenn Smith's lab), we did
> some in-house experiments and found it does help with problematic
> extracts that seem to inhibit enzyme (Taq) activity due to coextracted
> compounds. Since it's no too expensive, we just add it to all our aDNA
> reactions (unless we're working with ancient bovids, in which case we
> use Rabbit Serum Albumin). We haven't had a problem with it becoming
> contaminated, although in my lab our stock is aliquoted into very small
> volumes. But if the consensus in the UK is that it's a waste of time
> and money, I'd love to see some research showing that.
>
> best,
>
> Rika
>
> Dr. Frederika Kaestle
> Departments of Anthropology and Biology
> Indiana University, Bloomington
> USA
>
> Susannah Baldry wrote:
> > Dear Odile,
> >
> > I am not sure which protocols in the past used BSA in the PCR mix, but
> > (after throwing your question at my office colleagues) we agree it is
> > not common now - except for some commercial enzyme kits for arrays and
> > sequencing. In the latter case you would be following manufacturer's
> > protocols which would be exhaustive. If a published paper specifies
> > the contents of the PCR reaction it should be complete & repeatable,
> > so you will be safe following their protocol as described.
> >
> > Good luck with your experiments,
> >
> > Kind regards,
> >
> > Susannah Baldry
> >
> >
> > Odile Loreille wrote:
> >> Hi everyone,
> >>
> >> I'm back with a second question. Reading the latest aDNA papers, I
> >> noticed that many PCR protocols don't mention BSA anymore.
> >> I was wondering if you, aDNA specialists, omitted this information
> >> because you believed that the presence and amount of BSA isn't worth
> >> mentioning or if you truly stopped using it in your amplifications.
> >>
> >> If you want your response to be private, please simply write to
> >> [log in to unmask]
> >>
> >> Thank you very much
> >>
> >> Odile
> >>
> >
> >
> >
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