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uday

Member Since 19 Jul 2012
Offline Last Active Jun 08 2013 06:03 PM

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Active Posts 21
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My research interests
microbiology and molecular biology

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pcrman
26 May 2013 - 22:02
Maya1988
11 Dec 2012 - 07:11

Topics I've Started

High fidelity PCR trouble shooting

23 May 2013 - 06:59 PM

I want to amplify a 1.8kb gene using error prone PCR for subsequent cloning purpose. My primers works well with normal tag polymerase (Fermentas Dreamtaq Green PCR master mix) and gave me the desired product. However, when I use the same primers and template with high fidelity tag polymerase (Platinum Tag DNA polymerase high fidelity - Invitrogen), I can't get any amplified DNA. What was the wrong? what would need to be changed? I am bit confused

storage of culture plates

11 May 2013 - 07:45 PM

I am wondering why my culture plates (agar plates fresh or inoculated either with bacteria or yeasts) become filled with moisture when stored at refridgerator even though they are well wrapped with parafilm ? Anybody knows a solution ???

leaky promoter

21 April 2013 - 05:48 PM

What is a leaky promoter ? Is that refer to such promoter that can still do gene expression without the inducer ? Clarify what makes them so?

Molar Calculation

19 November 2012 - 11:15 AM

What a shame ! I got stuck in molar calculations.

Can anybody show how to do the math for making a 100mM, 100uL solution of X from the commercial available X solution of 96% ?

The details of the X are as follows.
96% assay
MW 222.37g/mol
density 0.879 g/ml

i.e. How much of commercially available X to be added ?

Low PCR product

14 November 2012 - 01:32 PM

i am getting a low PCR product ! I have tried reducing the annealing temp, increasing the primer and DNA template conc. too. But still quite low. Anything I should try ?