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Questions related to Cloning
We produce exogenous dsRNA in E. coli HT115 transformed with L4440 plasmid, but depending on the construct, we obtain different amounts of dsRNA. On what it can depend?
28 September 2022 9,167 0 View
Does anyone know whether the tracrRNA, Cas9 and the CRISPR locus in the CRISPR-Cas9 system belone to the same operon? Thanks in advance!
11 August 2022 9,217 0 View
We analyzed gene expression of A549 cell lines that were cultured in different conditions. The fold change data results of B-Actin as a housekeeping gene is significantly different from fold...
11 August 2022 1,545 5 View
As a part of my research work, I need to perform western blotting to detect the MDM2 levels in MCF- 7 cells. For this purpose, I need to purchase the antibody. Can I opt for monoclonal antibody?...
10 August 2022 2,878 0 View
As an alternative to the usual IP approach of tagging a protein in cells (trypanosomes) and passing the lysate through affinity columns, which has been time-consuming and unsuccessful for my...
06 August 2022 4,324 6 View
Is Ser/Thr protein kinase transmembrane protein in S.aureus ?
27 July 2022 4,872 2 View
image analysis question
20 July 2022 1,856 0 View
May I know if anyone have experience in culturing UC-MSC into organoids using Matrigel. I have dissected Mouse lung, dissociated them into single cells and added Matrigel, hoping that it will...
19 July 2022 6,582 1 View
Hi, I am trying to perform KD of my protein using lentiviral inducible shRNA, am working with U2OS, A549 and MDAMB231 cell lines. As a viral packaging control I used non-inducible lentiGFP, which...
12 June 2022 7,399 0 View
Why is my protein ladder getting smudged after transfer? I am transferring at 350 mAmp RT for 2 hours.
09 June 2022 1,095 2 View
The primers designed should be allele specific so as to amplify either my WT or Mutant transcript and not both.
01 June 2022 6,497 6 View
I have identified and characterized the new phage I have found some months ago, and I was able to capture the electron microscopy pictures and also sequence it. But I have had a serious issue...
25 May 2022 1,092 3 View
Hi all, here I have doubt on interpretation of #wiberg_bond_indices, I have Ni (coordination compound ), Ca(ionic compounds) complexes WBI values . Imp : In the above compounds both ligands are...
14 May 2022 4,116 0 View
Hi, I am a newbie with Hybridoma fusion and selection via ELISA and would be greatful for some tips for a screening problem I have. I completed the fusion successfully last week and there were...
14 May 2022 385 3 View
I would like to use the SKBR3 cell line in a long term co-culture with engineered T cells. To reduce their proliferative capacity and prevent rapid depletion of the media I would like to irradiate...
07 April 2022 5,510 0 View
Hello everyone, I am trying to overexpress a protein into BL21DE3 pLysS and have successfully cloned the gene in DH5α and checked the plasmid via double digestion to check the fallout. The...
06 April 2022 3,253 8 View
Why the changing of variance of the response variable (heteroscedasticity) is important for a GLM model e.g. Poisson? Why not just fitting an exponential curve?
18 March 2022 2,329 3 View
It is a MDA-MB-231 cell culture and we do not think in contamination because there is not movement or turbidity
15 March 2022 3,814 7 View
Hi, I am trying to express a gene which belong to classical TA (Toxin-Antitoxin) module. Specifically, I am trying to express the toxin gene using pBAD vector in BL21 cells. Despite trying out...
09 March 2022 8,251 2 View
I'm prepping drosophila ovaries for immunofluorescence. The protocol I'm using uses RNAse A and Propidium iodide for the nuclear stain. My understanding is that if I use DAPI I don't need a RNAse...
01 March 2022 2,794 3 View
Has anybody carried out cloning of viral genome using phi 29 DNA polymerase? If so, can you suggest me what are the components required for the process?
02 February 2022 1,108 0 View
I am working on virus and I want to clone the viral genome into a plant for the purpose of screening. Rolling circle amplification (RCA) is one of the methods for cloning viral genome into a...
01 February 2022 7,171 0 View
There are numerous articles on this topic already by google. Just to wonder are there some tools that are frequently used by biologists and can be related to experimental measurement?
01 February 2022 7,352 2 View
I know phi 29 DNA polymerase is sued to amplify the viral genome. Can the Pfu DNA polymerase be used to amplify the viral genome instead of phi 29 DNA polymerase?
01 February 2022 7,006 3 View