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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....

Read story →    11 comments    ****-

Differences between MSP and BSP methods

Hello everyone I'm new here and I was wondering if I could get some assistance. In my research of the literature I thought the differences between the two methods were based on the proximity of the primers to the CpG sites and that BSP primers...

Read story →    4 comments    -----

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...

Read story →    6 comments    -----

Issues Might Meet in the Process of Using ELISA...

Jul 03 2013 04:00 PM | dynah in ELISA

There are some issues that researchers might meet in the process of using ELISA Kits:

1. Positive results in negative control

2. High background across entire plate

3. Low absorbance values

4. High absorbance values for samples and/or positive control - absorbance does not go down as the sample is diluted down the plate

5. Inconsistent absorbance across the plate

6. Color develops slowly

Read story →    5 comments    -----    Elisa Kits, Creative Biomart

How to convert g into rpm?

May 28 2009 07:05 PM | bioforum in Centrifugation

The relationship between revolutions per minute (RPM) and relative centrifugal force (xg) is: g = (1.118 10-5) R S2 where g is the relative centrifugal force, R is the radius of the rotor in centimeters, and S is the speed of the centrifuge in revolu...

Read story →    4 comments    -----

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 →    3 comments    -----    RNeasy, contamination, RNA and 1 more...

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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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...

Read story →    6 comments    *****    promoter, tss, sequence, database and 1 more...

How to get rid of PCR primer dimer

Jan 29 2005 08:06 PM | bioforum in RT-PCR

1) Try the DMSO up to 5%. Some polymerases are ok with DMSO and others aren't. You never know until you try. 2) Try a two step PCR. There is a paper in Biotechniques about a two step PCR reaction but I don't have the reference with me sin...

Read story →    8 comments    -----

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 →    6 comments    -----    real time PCR, house keeping gene and 2 more...

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