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Questions related to Molecular Biological Techniques
I do not have a plasmid containing the NLS. I was planning to add the NLS to the sequence of a primer. My question is: Do I just place the sequence straight after the Start codon without a linker...
30 August 2016 831 4 View
by using the overlay method of antagonism the results are good and there is a clear zone but with ethyl acetate extract there is no results.
27 August 2016 6,430 5 View
I am totally confused about the RNA agarose gel. I have isolated total RNA from S. aureus. I want to see it in agarose gel. I ran 1% gel with formaldehyde and added 1 ug/ml EtBr during casting....
26 August 2016 8,047 7 View
I am looking for a reagent that makes the lysis of blood cells in order to improve the yield of DNA recovered by the use of total DNA extraction kit. Does anybody know this reagent ?
25 August 2016 2,777 1 View
Hi, I am studying genes for the resistance to the antibiotics in Klebsiella and I am analysing an outbreak of carbapenem-resistant isolates. The gene I am analysing is OXA-48 and it is known to...
23 August 2016 873 3 View
Hi, what is the best protocol to isolate DNA from FFPE tissue. I have tried KAPA Express Kit, NucliSENS Easymag, Organic Extraction, please advice further. Thank you!
23 August 2016 7,733 6 View
I got >2.0 values and I can't send it for NGS due to protein or humic acids contaminations I think. Thanks
23 August 2016 4,833 12 View
Hello everyone, I try to purify a High molecular weight protein. But i have a big problem because it forms aggregates. I try to add glycerol (5-10%) or increasing the NaCl concentration of 0,15M...
23 August 2016 4,228 10 View
The stratagene bacterial match II two hydrid system uses M9 ammonium as base media for growth. The recipient strain has a hisB mutation which is complemented with yeast his3 gene upon...
19 August 2016 3,074 3 View
I want to do research to compare mRNA in two groups and want to calculate the sample size for the study. Most of the articles has given expression of mRNA as fold change. Please help me how can i...
19 August 2016 1,295 4 View
Looking to experiment ways to speed up the process of protein digestion for nucleic acid purification using Proteinase K. Currently our lysis buffer includes low concentration SDS and EDTA but not...
19 August 2016 4,337 6 View
The culture looks fine during maintenance but after transfection, precipitation forms immediately after I adding the antibiotics (hygromycin b) for positive cells selection. So I tested the fresh...
19 August 2016 3,024 3 View
I am trying to amplify a large DNA fragment (about 5 Kb) by PCR. I used a FX-PCR master mix (for long and fidelity PCR). The PCR condition is as follow: 95C 5 min, 35 cycles of ; 95C 30 sec, 53C...
19 August 2016 9,851 14 View
Our lab is looking for some voltage gated sodium channel antibodies. Please write me if you know someone or a company who would have a good antibody. it will be a great help. Thank you -Gurjot
18 August 2016 8,184 3 View
I am using puromycin at 0.5 ug / ml in my cells, INS1. Although all cells are GFP positive, the level of my knockdown is around 40 - 60%. If I increase the puromycin concentration, that would...
18 August 2016 8,389 7 View
I am preparing ChIP-seq libraries for Illumina multiplexing. I have large amount of the ChIP-input DNA compared to the ChIP-output. I wonder if it is better to keep the DNA amount for library...
17 August 2016 1,160 4 View
Hi, I am doing semi-nested PCR. After 1. round with outer primers I can see specific fragment, but for higher sensivity I want to carry out nested PCR. Unfortunately, inner primer doesn’t work....
15 August 2016 6,381 4 View
I need to send Fosmid DNA for sequencing, but they told me the concentrations were too low & there are multiple bands. I tried mini-prep Qiagen kit many times & also midi-prep. How can...
14 August 2016 5,502 2 View
I used a single restriction enzyme, so the first problem was likely religation of the same plasmid. I treated with 10 units of CIP (about 5X overage) for only 5 minutes at 37C, and now there are...
14 August 2016 8,347 13 View
Sometimes, after plating transformed bacterial cells, colonies grow on plate are of different diameters. What are the factors responsible for uneven sizes of colonies on plate?
13 August 2016 1,022 5 View
I am identifying Paecilomyces variotii isolates through a poliphasic approach. At the molecular side, I have extracted the fungal DNA from culture in YES using a protocol with PrepMan (Life). The...
12 August 2016 1,789 1 View
Hello all. Recently I had an issue with a plasmid concentration after a mini-prep being recorded higher than it appears to be in experiments. That is not the nature of the question however; it is...
11 August 2016 4,562 3 View
I want to find new protein target for a compound with several -OH residues. Thus, I immobilized the compound on CNBr sepharose beads (GE Healthcare Life Sciences), following their protocol for...
09 August 2016 7,155 3 View
how do i quantify dna using nanodrop?
09 August 2016 4,790 5 View