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tech123

Member Since 26 Jul 2012
Offline Last Active Dec 04 2012 11:35 AM
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Topics I've Started

Adding T7 promoter sequence to vector

29 November 2012 - 08:54 AM

Hi all:
I am trying to make RNA probe for ISH. I ordered cDNA clone from a company and I realized that the vector they used does not have T7 or T3 or SP3 promoter sequence. So, how can I add the promoter sequence to my vector? Also, I streaked the glycerol stock which I got yesterday but no colonies today, what could have gone wrong?
Vector:pME18S-FL3
Thanks in advance

Why can't we detect multiple protein in a single membrane using western blot

07 August 2012 - 09:17 AM

Hello:
I am trying to detect three proteins A, B and C. Unlike Immunohistochemistry, Why i cannot detect all the proteins in a single membrane when the antibodies against my protein are raised in different animals? I know the size of my proteins.
Thanks

Wet transfer: western blot

26 July 2012 - 02:49 PM

Hi..This is my first time doing wet protein transfer. I use PVDF membrane. Run at 70 V for ~ 3 hrs. After transfer my membrane becomes little dry and the outer coat of the membrane starts to come out leaving dark spots on the membrane. Also do protein ladder go bad? They have started to appear very dim in gel. Any suggestions!!
Thank you

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