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leyshmanioz

Member Since 01 Apr 2009
Offline Last Active Jan 27 2013 12:33 AM
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Topics I've Started

Amplification of CAG promoter problems

26 January 2013 - 11:04 PM

Dear all,
I have problem amplifying CAG promoter, apparently its very GC rich some regions have 70% . We've tryied Phusion HS from thermo, Turbo polymerase from Agilent , Taq from NEB, tried different DMSO concentrations, different primers from 55 to 66 C, GC rich buffer, MgCl2 concentrations.
Did anyone have tried to PCR amplify this promoter, would appreciate the help.
Thank you

Which isotype control should i choose

14 January 2013 - 11:04 PM

Dear all please help  a FACS noob,
I need to choose isotype control for my surface marker staining. Should i choose isotype control depending on the species i'm labeling, so if i have Ab against human CD44-PE conjugated my control should also be anti human IgG. Also does the choice of type of Ab depend on the specific stained one. For example if my Cd44-PE was IgGk then isotype control should also be IgGk and what is the difference between normal IgG, does it really make a big difference.

Thanks

mean fluorescence intensity, ratio

31 December 2012 - 08:11 PM

I am confused in flow data analysis, Can someone explain me what ratio of MFI show us. meaning GFP/RFP ratio.
Thanks

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