Messenger Affinity Paper

Keith Hutchison KEITHH at MAINE.MAINE.EDU
Wed Jul 10 09:32:59 EST 1991


In article <0094B4BF.4A5AE080 at m44.unm.edu>, kim at m44.unm.edu says:
>
>I have been looking into a method for Poly(A+) selection using a paper
>substrate linked to poly(U) called Hybond-mAP (AMERSHAM).  I am told (by
>AMERSHAM technical support) that mAP has a small but loyal following, and I
>was
>wondering if anyone out there has used it.  Is it as easy and effective as
>advertised?
>          Thanks in advance for your comments.
>Daniel Kim


I guess we are among the loyal users of mAP.  We find that it gives us
comparable yield and quality to oligo-dT with greater ease of use.  We
have made one minor modification which we found was necessary to give us
adequate yield.  We pre-incubate the filter in the RNA solution for 5
min at RT before blotting the RNA as given by the supplier (Plant
Physiol 94: 1308).  The mAP also has the advantage in dealing with
samples that are highly viscous due to carbohydrate (some of our samples
have enough to choke a horse let alone an oligo-dT column).  In this
case simple incubation of the filter in the solution will retrieve
enough poly-A RNA for libraries or blotting.

   Keith Hutchison
|==========================|==================================|
| Dept. of Biochemistry, Microbiology and  Molecular Biology  |
| The University of Maine, Orono, ME  04469                   |
| BitNet: keithh at maine    Internet: keithh at maine.maine.edu    |
| Phone:  207-581-2827                                        |
|=============================================================|



More information about the Methods mailing list