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Questions related to Molecular Biological Techniques
Hi everyone, A while back a lithium acetate transformation of a triple knock out mutant Candida albicans strain was performed yielding several transformants. The transformants, however,...
07 April 2019 4,352 5 View
I am having problem obtaining the desired confluency. They are not growing out well even after 24-72 h incubation. Confluency remains less than 60% Note: - Media is RMPI 1640 (As recommended by...
16 January 2018 1,011 4 View
It should be possible to induce germination also on slides or agar plates. I have already tried different methods: aqueous solution, film of water on slide, nutrient solution with dextrose,...
02 November 2017 4,848 2 View
The protein I want to express do have Zinc binding site, it actually use Zinc as a cofactor. In this case, do I need to add ZnCl? Ps, the bacteria strain I'm using is Rosetta.
02 November 2017 6,398 7 View
Will the magnetic beads (SIGMA-ALORICH; ANTI-FLAG M2 Magnetic Beads; Catalog Number:M8823) that suspended in 50% glycerol be freezed at -20 ℃? In my oponion, stored in glycerol is to avoid freeze,...
31 October 2017 9,334 4 View
I recently did an experiment where I created sterile media by adding 1mM of ammonium sulphate to Murashige and Skoog (MS) Modified Basal Salt Mixture (No Nitrogen) M531. The media was not buffered...
31 October 2017 3,682 4 View
Hi! I would like to discriminate dead cells from my population using LIVEDEAD fixable dye. In principle it stains amine residues in both live and dead cells, but because of cell membrane...
29 October 2017 5,244 3 View
Hello. We recently bought Cruz Marker Molecular Weight Standards from Santa Cruz Biotech and for some reason, we only see one band (we think is the 90KDa, but since we can not see the others...)...
07 August 2017 2,815 7 View
Hello, I recently got 4-20% gradient gels from Biorad to try for better detection of 4EBP-1 protein in horse muscle tissue. But I am running into trouble during the electrophoresis step where...
01 August 2017 6,144 3 View
Hi everyone, We see two transparent lines (like the one separating the stacking and running gel) during the running and we also note that 25 and 20 MW band get stack in the upper transparent band....
31 July 2017 3,274 5 View
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 5,236 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 4,977 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 2,951 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 659 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 8,421 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 3,289 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 8,541 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 3,112 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 7,562 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 529 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 7,378 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 3,348 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 2,876 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 3,707 5 View