internal standards for northerns

J. Hendriks 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
>     ratio....
>
>     deb
>
>
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.
Brenda Hendriks
e-mail: b.hendriks at wkz.azu.nl or hendrikb at worldonline.nl





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