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Questions related to Molecular Biological Techniques
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 1,310 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 4,353 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 3,042 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 9,929 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 8,197 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 8,412 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 1,177 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 6,390 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 5,523 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 8,359 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 1,045 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 1,803 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 4,576 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 7,178 3 View
how do i quantify dna using nanodrop?
09 August 2016 4,812 5 View
Planning to express two sets of genes separated by 17 nucleotides. Have upstrem sequence but really dont know whether the two genes have same promoter or different ? I expect that they will...
08 August 2016 2,403 2 View
Dear All Hi, I am writing to ask your opinion about chance of success designed real time Taqman probe as: 5'- TAC GCC GTA YCT AGC TWY YGC CTG GAC CGT C -3’ as you see in about middle of probe...
08 August 2016 9,561 2 View
I have Medroxyprogesterone acetate in powder form which I want to dissolve it in DMSO but, I am not sure what concentration of DMSO I require.
08 August 2016 6,889 3 View
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07 August 2016 5,905 5 View
I used a TOPO TA kit which contained a linearized vector of 3.9kb to ligate my DNA of about 3.8kb. After transformation and plating on LB agar containing X-GAL I got quite a number of white...
07 August 2016 9,911 3 View
Specifically 25 uM Sigma I2764 ML-7 in DMEM low at 37 degrees C.
04 August 2016 6,259 1 View
in-fusion cloning is In-Fusion® HD Cloning Kit by Clontech® Laboratories
04 August 2016 468 2 View
i am using stripping buffer for 1hr to remove the previous bands but still the previous band are visible. is it possible to completely remove the previous bands with stripping buffer ? .
04 August 2016 2,498 3 View
I need isolate the CDV in cell culture.
03 August 2016 2,436 3 View