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Questions related to Molecular Biological Techniques
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 3,385 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 2,903 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 3,747 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 2,188 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 323 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 7,873 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 7,799 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 7,341 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 9,486 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 2,656 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 9,631 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 5,757 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 5,039 3 View
I prepared a mastermix for technical triplicates for SYBR green assay (reaction volume 10ul) Out of the triplicates, usually the 3rd replicate will have difference in Ct value i.e. later Ct (range...
11 February 2017 3,279 3 View
I want to clone a 7 kbp pDNA with a 1.2 kbp fragment and I have already tested the 1: 3, 1: 1 and 3: 1 ratios but I always get self-ligation of the vector. How can I resolve this or improve the...
11 February 2017 2,339 4 View
I ran samples (looking for c.bovis in swabs and tumors) using the same primes/probes, Taqman Master Mix, DEPC H2O that was used last week on samples that worked fine. These new samples that should...
10 February 2017 2,875 4 View
I want to construct a vector to drive shRNA expression by U6 promoter. However, due to limited genetic space I would like to use minimum possible nucleotides of U6 promoter. Do anybody know...
10 February 2017 5,010 2 View
Hi, I have a project about using the micropatterns substrate to see the hMSCs behavior and the differentiation fate. the project goals is to find out whether the micropatterning can lead the...
08 February 2017 3,697 3 View
I'm having a hard time finding a reference for my specific conditions, any help would be appreciated: I have dsDNA bound to streptavidin-coated magnetic beads via a biotin interaction. The dsDNA...
06 February 2017 1,654 1 View
Hi everyone, I need a solution for an issue that I have with my AFLP protocol.Currently, I've been trying to make some AFLP's from a grass species that I study for my research project. However,...
30 January 2017 9,739 3 View
i did cloning for cytochrome P450- 2D6 into pGEX-6P-2 plasmid vector. when i screen using PCR i get the band of PCR product at an actual size of 1375 bp. but when i use double digest i do get...
30 January 2017 4,768 3 View
I have amplified 500 bp up- and downstream of my target gene and placed a KanR for selection between both 500bp fragments. Is that correct? Now, I want to use pGMT-t vector (Promega) as a...
27 January 2017 8,668 3 View
When determining titer of T7 phage by plaque assay, I get a gradient of plaques where one side of the plate is clearing and the other half of the plate is a lawn of bacteria. Anyone experience...
24 January 2017 276 4 View
I used Qiagen RNeasy Plant Mini Kit to isolate RNA from Turmeric Rhizome (6 months old) and used tertiary rhizomes as a sample for RNA isolation. I used two varieties (First Three columns...
21 January 2017 7,727 13 View