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Pinned  How to avoid contamination in PCR?

Oct 02 2004 12:24 PM | bioforum in PCR

PCR contaminations are like ghosts... you know they are there but you dont know WHERE?? I have followed the following with good results... Prepare your master mix in a separate room from your current location, somewhere PCR is not the main practice....

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Useful Dilution Techniques & Calculations

Mar 21 2009 02:33 PM | bioforum in General Lab Techniques

Today was a very bad day at the university... beside that some students answered in the quiz question that if you get blue color in Gram staining that means your bacteria is Gram negative :huh: I noticed most of the students including myself had troub...

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How to coat coverslips before seeding cells?

Aug 18 2011 05:55 AM | jennycharlies in Cell Culture

jennycharlies Posted 18 August 2011 - 06:55 AM Hi, I am going to seed my cells onto coverslips inside the wells of a 12 well plate and was wondering what is the best way to coat the coverslips? I have 18mm coverslips and need to coat with poly-D-...

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Tips on freezing and thawing of cells

Sep 10 2004 12:32 PM | bioforum in Cell Culture

I am facing problems in thawing of cells. The procedure which we follow is- we take stock from liq. N2 then though it in water bath at 37 degree. Then we take medium in falcon (5ml) then add thawed cells dropwise into it. We then centrifuge it at 1500...

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How to find promoter sequence for methylation s...

Aug 05 2009 07:43 PM | pcrman in Bioinformatics and Biostatistics

Many people have problem identifying or predicting the promoter sequence of a gene, or don't know how to get the actual sequence for analysis such as primer design, transcription factor binding site search, etc. Here I provide ways how I do these t...

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DNA migrate differently in agarose vs PAGE gel

Aug 15 2011 08:59 PM | pcrman in Electrophoresis

i am currently working a gene in leukemia. The mutant allele has been sequenced to contain 527 base pair. when i run an agarose gel to confirm, it did appear in the position approximately equal to 527 base pair according to the DNA ladder. HOWEVER, whe...

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Can 18s gene be used as reference gene for RT-PCR?

Mar 10 2013 06:31 PM | BMF in RT-PCR

Hi, I have done few real time PCR works recently, and I used the 18s gene for endogeneous control. Its Ct value was approximately 11 for every cDNAs used as template. Here, I have a question. Since 18s gene doesn't have poly A tail, it can't...

Read story →    4 comments    -----    real time PCR, house keeping gene and 2 more...


Normalization for RNA or cDNA during two step R...

Apr 20 2009 09:13 AM | bioforum in General Lab Techniques

Curtis 20 April 2009 - 03:13 AM Hello all, I have a two-step SYBR Green RT-PCR kit for RotorGene. After I extract RNA from my samples, I proceed with cDNA synthesis right away without measuring the RNA concentration or wit...

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Common causes for low RNA A260/230 ratios

Jul 30 2013 01:38 PM | robradford in RNA Methods

Discussion on things that cause a low A260/230 ratio. Please add your comments.

Read story →    1 comments    -----    RNeasy, contamination, RNA and 1 more...


How to avoid foaming during sonication in ChIP...

Feb 01 2009 01:33 PM | bioforum in ChIP Assay

ChIP - Foaming in sonication and other woes - <small> (Jan/27/2005 )</small>Hi all! I was wondering if anyone out there has got any trick to avoid foaming during sonication in the SDS lysis buffer for ChIP assays. Supossedly, placi...

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