19 Questions 64 Answers 0 Followers
Questions related from Maynak Chakraborty
I am a beginner of Transcription factor ChIP experiment. I have taken 30ug of protein for each immunoprecipitation and the input was 1%. After immunoprecipitation, my input DNA concentration is...
02 February 2020 1,903 2 View
I am planning to do an analysis of both allelic and genotypic level of association of an particular SNP with a disease via case control study.But, I can not understand which statistical test...
08 August 2019 8,267 3 View
For my promoter activity analysis, I take 0.12pmol of each different size of construct and 1:100 ratio of renilla vector. I balance the amount of total vector using a filler plasmid. 1) Construct...
03 March 2019 6,535 2 View
I am doing site-directed mutagenesis using Inverse-PCR method to mutate a 10bp region of my insert. My vector plus insert size is 5.6kb. I am using Pfu polymerase from NEB to amplify my plasmid...
07 July 2018 2,235 5 View
I am doing Dual luciferase assay in a hard to transfect cell line and got these values- 1) Non-transfected- Firefly 93.16(RLU) and Renilla 123.85(RLU), so F/R is 0.75 2) Empty vector...
05 May 2018 2,064 2 View
I want to check the activity and the efficacy of 5 different drugs in cell lines to evaluate the best one based on their potency and reduced toxic effects. In many papers, researches did take the...
11 November 2017 8,520 11 View
I am a beginner of cell culture experiments and just have started some experiments with Hela cell line.In research papers, it is always highlighted that, each experiment has been done in...
08 August 2017 7,264 2 View
In my cell culture experiments, first I have revived a vial of cells, subcultured it, then extracted RNA from a particular no of cells for my experiments.After RNA isolation, I have freezed my...
07 July 2017 8,466 1 View
I am a beginner of western blot and my target protein if 124kDA whereas the endogeneous controls are 41kDA and 55kDA. I did wet transfer at 100V, 80mins and successfully got the band for target...
06 June 2017 4,700 5 View
05 May 2017 9,210 10 View
I am a beginner of western blot and from some days I have been facing a lot of problems . My experimental protein size is 124kDA and I choose Beta Actin as a loading control which is small...
04 April 2017 1,831 7 View
I am interested to understand the genetic pathophysiology of Fuchs Dystrophy and for that purpose it is required a cell culture based model system of Fuchs corneal endothelium cell line. We have...
03 March 2017 3,947 4 View
I want to amplify a 3kb gene and its four fragments using PCR. Using Taq DNA Polymerase and Taq buffer I have already standardized the Ta of the primers ( whole gene and its fragments). Because I...
11 November 2016 2,988 7 View
For luciferase assay I want to clone my gene using Kpn1 and Nhe1 as both of these are present in vector but not insert.My clone of interest is human genomic DNA.I have designed my PCR primers...
11 November 2016 4,900 8 View
I have done competent cell preparation for cloning ( DH5 alpha) by Rubidium chloride method first time.But, by mistake the entire experiment I have accomplished outside the hood.Is it okay or I...
11 November 2016 6,613 7 View
I want to make cDNA of my gene with two restriction sites at its ends for cloning and over-expression studies.I found that most of cDNA , commercially available are GFP tagged. I am little bit...
10 October 2016 7,126 3 View
I want to know whether transcription factor X binds to promoter region of my gene or not through luciferase assay and like to prefer a vector which has minimal promoter plus MCS site upstream of...
10 October 2016 3,800 6 View
I want to create a knock-down, stable cell line using crispr-cas system without using viral based vectors. I want to regulate the knock-down intensity around 60% so that I can do my experiments...
09 September 2016 10,056 3 View
I want to knock-down a particular gene and try to generate a stable cell line for observing the effects. I can not knock it out because it has a significant role in cell proliferation. How can I...
09 September 2016 5,564 2 View
