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RNase

A transferase that catalyzes the hydrolysis of ribonucleic acid.


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RT PCR contamination.

... controls, a -RT and a -template control, both of which are firing. I have changed everything (New; DTT, dNTPs, Primers, water{Sterile MQ}, RNase out, buffer) and have not found the contamination, I have confirm the negatives are firing by endpoint PCR with primers designed to span an intron ...

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by georgewiggins
Thu Sep 06, 2012 9:49 pm
 
Forum: Molecular Biology
Topic: RT PCR contamination.
Replies: 2
Views: 4161

PCR from cDNA question

... RNA was extracted from ~100 whole embryos (~1000 cells each) of an invertebrate marine species using QIAGEN RNeasy Plus mini kit eluted with 40ul RNase free water. Nanodrop gives a low reading and a weak absorption peak (~10ng/ul). Gel analysis of 3ul such RNA yielded no bands (not even the ribosomal ...

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by vastgenome
Fri Jun 22, 2012 3:55 pm
 
Forum: Molecular Biology
Topic: PCR from cDNA question
Replies: 1
Views: 3230

Can I freeze RNA that is atteched to a column?

Cool, thanks for letting us know :) Btw, did you freeze them with some cryopreservant / RNAse inhibitor, or just as they were?

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by biohazard
Wed Nov 16, 2011 7:47 am
 
Forum: Molecular Biology
Topic: Can I freeze RNA that is atteched to a column?
Replies: 7
Views: 3521

DNA Extraction/Isolation

I've never heated TE buffer with DNA. What step are you talking about? You're adding some enzyme, right? Like RNase. Both heating steps serve to denature and inactive present enzymes.

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by JackBean
Fri Jul 01, 2011 3:08 pm
 
Forum: General Discussion
Topic: DNA Extraction/Isolation
Replies: 2
Views: 3319

Re: How do you overexpress and knock down a gene

... you can either express an antisense transcript or an shRNA, but these knockdowns will not be very specific. The antisense transcript will trigger RNase-H activity and the shRNA will use argonaute activity; both of these enzymatic systems will recognize mRNA with as little as 8 bases of complementarity. ...

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by jonmoulton
Tue Nov 30, 2010 10:27 pm
 
Forum: Molecular Biology
Topic: How do you overexpress and knock down a gene
Replies: 2
Views: 13567
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