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Questions related to Biochemical Techniques
I am about to start some new experiments that rely on TAT-Cre (from Millipore) to delete a gene of interest in cell culture in order to study the impact on cell differentiation and gene...
05 October 2014 6,466 5 View
I have been trying to generate clones using pENTR DTOPO for gateway cloning. I am able to amplify my insert, set up a reaction and get colonies. But when I do a colony PCR with insert specific...
04 October 2014 2,327 7 View
What is the optimum voltage and ampres? Thank you all in advance.
03 October 2014 1,028 3 View
I wanted to use proteinase K to treat conditioned media in order to determine if a protein factor is responsible for the effect seen when CM is used on cells. I have come across various protocols...
03 October 2014 4,226 3 View
Dear all. I'm trying to sequence the coding regions of a gene called NDST1 in a 400 individuals - population ( The samples of the population were collected in total DNA form). Problem is the gene...
01 October 2014 9,681 10 View
I want to extract genomic DNA from tumour cells. Can anyone help me how? most simple without PCI.
29 September 2014 7,866 4 View
Please give an explanation for the above question.
25 September 2014 9,239 4 View
I add filtered L-cysteine in cool anaerobic broth as a reducing agent. Is it possible to add L-cysteine in the culture media before autoclaving without compromising the reducing property of...
19 September 2014 3,836 26 View
I purchased poly dA-dT.poly dA-dT 10 unit packs from sigma and the concentration was found spectrophotometrically. I got a weight of this product in ug/ ml but I would like to prepare a uM...
15 September 2014 2,379 5 View
09 September 2014 1,766 5 View
When I dissolve DPPH in acidified methanol(15% 1N HCl) it is showing yellow color. what might be the problem, is it acid or methanol or DPPH?
09 September 2014 9,692 7 View
What is the chemical name of ZEN.
09 September 2014 6,758 1 View
Hi, I have recently encountered a loading problem with one of my blots. The data just didnt make any sense. I reprobed for actin. The loading problem should be correctable for actin normalization...
09 September 2014 6,363 4 View
I am looking for optimised protocol for western blot of 2D gels! any one routinely doing please help me!! I am missing minute details like voltage , buffer etc. Working wirth semidry GE...
09 September 2014 9,091 3 View
During the experiment and literature I found that many peoples had used tris Hcl for protein extraion in the range of 20 to 100 mM concentration. why peoples are not extracting proteins at...
09 September 2014 1,975 4 View
I am planning to isolate Islets from a rat pancreas using Ficoll density gradient of 1.108g/mL,; 1.096g/ML; 1.069g/mLand 1.037g/mL. I have bought Ficoll 400 powder and I would like to make these...
09 September 2014 1,163 3 View
I have problems determining the reliable dilution for my qPCR. I have repeated the experiment 3 times, it seems that I have gotten different results for every experiment. However by comparing the...
09 September 2014 3,073 4 View
Hi，guys！Does anyone know the difference between 2X and 5X sds loading buffer? It is said that maybe you cannot get appropriate results using 5X Loading buffer. Does anyone know the...
09 September 2014 6,353 14 View
09 September 2014 4,430 5 View
I wish to perform a chemical crosslink experiment on a homodimer protein with low thermal stability. Due to thermal stability issues, the desired crosslink needs to be done
09 September 2014 8,229 3 View
What is the optimal protein concentration of cell lysate used for this assay kit?
09 September 2014 8,835 3 View
09 September 2014 8,727 27 View
I am doing work on the isolation of antioxidant compounds. The new compounds exhibited higher antioxidant than the fractions in beta carotene bleaching but weaker in DPPH assay. Is there any...
09 September 2014 6,820 4 View
09 September 2014 2,422 6 View