internal standards for northerns
hendrikb at worldonline.nl
Mon Nov 10 14:37:57 EST 1997
deborah.hailstones at smtpgwy.agric.nsw.gov.au wrote in article
<9711078788.AA878855167 at smtpgwy.agric.nsw.gov.au>...
> In article <34512732.238CDC58 at facstaff.wisc.edu>,
> pemozdzi at facstaff.wisc.edu wrote:
> > What are some of the disadvantages of using an 18S or 28S probe as
> an > internal standard for a Northern? Are they better than GAPDH?
> I have routinely used 18S for standardisation, and to overcome the
> problem of signal strength and also to ensure that the probe isn't
> limiting (which would totally destroy any chance of quantitating using
> the probe...), i always add a bucket load of cold 18S oligo to the
> hyb.... ie add end labelled and cold together, in say a 1/1000
we have always used a Histon H3 probe as a internal control when working
with samples of the frog (different ages). Another possibility is the use of
an actin-probe. Be sure that you have not treated the samples with a
substrate that also have some influence on the internal probe you use.
Good luck with the experiments.
e-mail: b.hendriks at wkz.azu.nl or hendrikb at worldonline.nl
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