Contact experts in Molecular Biological Techniques to get answers
1,768 views 3,173 posts
Questions related to Molecular Biological Techniques
I have been getting exponential curves in my negative PCR reactions. I’ve performed extraction controls (with no sample extracted) and No Template controls (on the PCR) and they are all clear....
06 July 2017 5,256 6 View
I am having negative results for my DNA/RNA extraction from stool samples on Nanodrop for Concentration, 260/280 and 260/230. This is the first time I'm having this problem and have tried to use...
18 June 2017 5,010 12 View
I ran RNA denaturing gel (1.2% agarose, 1X HEPES-EDTA buffer and 0.45M formaldehyde). Used 0.5 ug of RNA, 5 ul of loading dye (HEPES-EDTA, bromophenol blue) and 1ul of Etbr in the reaction. After...
13 June 2017 2,974 4 View
I'm confused by my cloning PCR results.I have used the TOPO TA Cloning kit. I got colonies after blue-white screening. I grew them in the LB liquid culture in presence of ampicillin drug and then...
03 June 2017 675 2 View
Dear Colleagues I am bout to explore the extravasation of tumours by in vitro modelling using a trans well (Boyden chamber) assay; I must confess this is completely new to me In essence, we are...
08 May 2017 8,441 3 View
Hello Every one, I am wondering whether southern blot can determine the mutated genes while they will be used as probes. Will it show result if the DNA fragment used as Probe has mismatches at...
04 May 2017 3,313 2 View
Hello, If I use a PCR plate for library prep and I need the products after each step in the protocol (in my case it's polyadenylation, RT and PCR), is it safe to open the plastic foil that covers...
26 April 2017 8,558 3 View
I want to evaluate the amount of internal cAMP levels. Unfortunately, I suspended my cells of desire in RIPA buffer. Is it possibly, despite the harsh buffer, to determine cAMP levels in an...
24 April 2017 3,132 8 View
Hello everyone, I am about to perform a turbidimetric assay of Tyrothricin and for the purpose of the assay I need to make equal concentrations of standard solution (Gramicidin) which is 1081...
19 April 2017 7,575 2 View
I work with zebrafish adults and larvae and am trying to characterise mutations from CRISPR and ENU mutants. I either extract the genome from entire larvae between 3 and 5dpf or adult fin clips....
02 April 2017 544 1 View
Having problems with a RT-PCR reaction that works fine with regular Taq but gives absolutely no product with Phusion Hot Start II from Thermo. I replaced my dNTP stock with new material, tried...
31 March 2017 7,393 5 View
Hello, I am seeking for a method for isolation of HIV RNA from plasma sample. I am doing NGS so the isolate has to be highly pure (without any human or bacterial DNA/RNA). I tried standard kits...
30 March 2017 3,366 3 View
Hello Today i want to ask about whether there is optimal condition of centrifugation (speed, time, and temperature) to obtain cell pellet. I have already searched many protocols, but i only can...
27 March 2017 2,890 4 View
My experimental gene has 5 different transcription start sites.From different web based tools, I searched that the predominance of these sites in different tissues but still now I have not get any...
26 March 2017 3,729 5 View
Hi!! I am trying to do radioactivity based invitro transcription assay using oligo dc tailed template (pGL2CMV) but facing some serious problem. At this moment I am trying to standardize the...
26 March 2017 2,174 2 View
Hi. I'm working on BV2 cells. I'm having problem in NO production after LPS induction. I'm using 10μg/ml of LPS concentration and incubate it for 24 hours. . Sometimes the production of NO will be...
20 March 2017 304 3 View
When I infected 293T cells and MS7 cells with Lentivirus they lost their monolayer property and started growing on top of each other. They are under puromycin selection. My labmate did Crisper-...
09 March 2017 7,842 1 View
I have a sample that is a mix of DNA (~100bp) and maltodextrin sugar with some processing. I need to show the DNA size via gel electrophoresis (GE), but the DNA remains smeared at the top of the...
06 March 2017 7,779 6 View
Hi, we sometimes get this weird stains on the agarose gels. We tried re-making TAE buffer and agarose from scratch (after some experiments, we were quite sure it was the buffer), we obtained a...
01 March 2017 7,329 5 View
Hi , For the past 2 weeks I have been struggling with something so simple. I have a plasmid ( vector is pcDNA4/TO with amp resistance). It has worked perfectly before and recently I was close to...
27 February 2017 9,469 5 View
I have transformed my cloned plasmid DNA into bacteria and cultured bacteria. Then, I extracted DNA from bacteria and run in 0.5% agarose gel. But I am not seeing any DNA band except ladder. (In...
24 February 2017 2,637 14 View
I've done a full cleaning on the GC/MS including replacing my moisture indicator trap and included a new bulk moisture trap, replacing the septum and inlet liner, replacing the column and cleaning...
23 February 2017 9,622 10 View
I am trying to determine the cellular localisation of a lncRNA in THP1 cells. To do this, I have fractionated cells so that I have Cytoplasmic, Nucleoplasmic and Chromatin Associated fractions. My...
16 February 2017 5,739 2 View
Hi, I would like to quantify global chromatin compaction using MNAse treatment followed by Comet assay. However, I don't know if Comet assay would sensitive enough to detect effect of drugs on...
13 February 2017 5,031 3 View