Fungal 28s amplification primers

M. Alejandra Mandel via methods%40net.bio.net (by mandel1 from email.arizona.edu)
Thu May 27 12:56:23 EST 2010


You could try Rytas Vilgalys' website.
It has all of the rDNA primers, subdivided into groups 28S, ITS, 18S. For 28S,
it says that most molecular systematics studies only utilize the first 600-900
bases from the LSU gene ....  We covered part of this using ITS1F and LR3.
Vilgalys describes how they amplify this region and what primers they use for
sequencing.

This is the web address:

http://www.biology.duke.edu/fungi/mycolab/primers.htm

Quoting Adroit A <cognizann from googlemail.com>:

> Dear Bionetters,
>
> I am trying to amplify the 28s rRNA region of a fungus. For this i already
> isolated the genomic DNA and amplified it with ITS-5 and ITS-4 primers. I
> got a PCR product of length approx. 700bp. However this amplified product
> covers the region between 18s-5.8s-28s. Could anyone please suggest me the
> universal primers that specifically amplify the 28s region alone?
>
> I also used ITS-1 and ITS-4 primers for amplification, however no
> amplification was observed. Please let me know the reason for this.
>
> Thanking you in advance.
>
> Thanks,
> Adroit
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Ale Mandel





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