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SadamCorneli

Member Since 28 Oct 2012
Offline Last Active Jan 11 2013 09:03 AM
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#144228 A good UV transilluminator (electrophoresis)

Posted Trof on 29 October 2012 - 08:25 AM

In my experience, for routine and comfort gel imaging (like udes every week, not once in two months) is much better to look for complete imaging solutions. Those UV transluminators  with detachable hood with a common digital camera attached on the top just sucks, really. It's difficult to focus, change expositin times, everyting is controled on computer and there is no live-preview. I had to use it few times when our CCD camera was broken and it was hell.

These things however are not that cheap, but chambers with  black-and-white CCD camera have focusing, zoom, exposition control and live preview give much more better images much easier and quicker way.

Also as mentioned, for gel extraction longer wavelength UV option may be beneficial. If you plan to cut the gels under UV look for chambers that allow that (i.e. UV on while doors are open), some may not.


#144225 A good UV transilluminator (electrophoresis)

Posted phage434 on 29 October 2012 - 06:29 AM

You might consider a blue light illuminator, rather than UV, which will limit DNA damage.


#144205 A good UV transilluminator (electrophoresis)

Posted bob1 on 28 October 2012 - 08:21 PM

Any brand should work fine, there isn't much difficulty with most transilluminators.  If you need to do a lot of DNA gel extractions, it is worth getting a less damaging long wavelength option on the transilluminator.

A hood is useful for taking pictures and avoiding unnecessary UV exposure, but it isn't absolutely necessary - kodak used to sell a camera and hand held hood that could be shifted on and off the trans.


#138424 PCR primer design - published primers trustable?

Posted pcrman on 27 July 2012 - 01:25 PM

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1) Should I not worry that I can't confirm the sequence and just trust that since the article has been published that it will work?
NO, Don't trust any primers published in papers. The primer sequences could be wrong either due to bad design, sequence identification, or errors in making the paper.

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2) Should I even worry with published articles and just design my own based off the updated NCBI RefSeq (mRNA)?
Yes, I would first try to design my own primers. Published qPCR primers from high quality papers proven to work may serve as references

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3) Is there a way to blast primers against the entire genomic sequence? (I would have to invert the primers sequence first, correct?)
Yes, use UCSC in silico PCR tool. But be cautious that if a primer span exon-exon junction, the tool may not return a hit.


#142011 Help with sequencing of a 120bp PCR product.

Posted bob1 on 21 September 2012 - 11:17 PM

You could also try denaturing HPLC.


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