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Dot blot

Dot blot

(Science: molecular biology, procedure) method for detecting a specific protein or message. A spot of solution is dotted onto nitrocellulose paper, a specific antibody or probe is allowed to bind and the presence of bound antibody/probe then shown by using a peroxidase coupled second antibody, as in western blotting or by other visualisation methods.


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Souther blot and restriction mapping

... restriction maps from fragment data obtained through endonuclease digestion and separation by standard gel electrophoresis, however, Southern Blot data only shows those fragments that a probe has hybridised to and I am struggling to find the correct approach. The parting words from lecturer ...

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by Mistersnuffle
Sat Oct 04, 2014 11:17 am
 
Forum: Genetics
Topic: Souther blot and restriction mapping
Replies: 0
Views: 25

mRNA vs. cell reporter assay

I have the same question, phrased in a slightly different way: Maybe the reporter system provides us with more information than RT-PCR/Western blot, because it proves that the transcription factor is actually active too, not just present? I thought it is enough to detect a significant increase ...

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by kk
Mon Sep 08, 2014 2:57 pm
 
Forum: Cell Biology
Topic: mRNA vs. cell reporter assay
Replies: 2
Views: 698

DNA testing times

... done in a working day, though the PCR thermal-cycling machine can be set up and left to run overnight. For most DNA methods, whether PCR or blot, an hour is much too short. The time required for analysis will depend on how much information the assay provides and how complex a question you ...

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by jonmoulton
Fri Apr 11, 2014 2:47 pm
 
Forum: Genetics
Topic: DNA testing times
Replies: 2
Views: 1214

Re: Western transfer failure

... in transfer buffer. I was transferring two 12% SDS-PAGE gels to PVDF in one tank. 100V, 1 hr. One gel transferred fine, another went backwards to blot paper after 1 hr. Did I just arrange backwards? Did lack of SDS mess with electric gradient? Correlation may not be causation, so I don't claim ...

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by yanggers
Thu Feb 06, 2014 9:16 pm
 
Forum: Molecular Biology
Topic: Western transfer failure
Replies: 5
Views: 5241

Re: qPCR vs Northern Blot

Thank you both for answers. Firstly - aliquots for Northern blot and qPCR were the same (firstly I did NB and after a couple of weeks qPCR). @jonmoulton - you're rigth first - in Northern blot I had silencing, in qPCR i didn't. As far as your question ...

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by Nevthonien
Tue Nov 12, 2013 2:47 pm
 
Forum: Molecular Biology
Topic: qPCR vs Northern Blot
Replies: 4
Views: 2789
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