Avinash Sharma

13 Questions 16 Answers 0 Followers

Questions related from Avinash Sharma

For confocal microscopy, is there a way to tag ubiquitin protein without hampering its job in vivo?

Hello everyone! I am planning an experiment where I would like to check ubiquitin transfer using confocal microscopy; thus, I intend to add a fluorophore tag to the ubiquitin molecule. I read a...

13 August 2023 7,492 1 View

Can you identify and tell me the term for these cells in Arabidopsis thaliana?

Please see the attached image for the reference. I have been seeing these cells in the Arabidopsis thaliana seed (post germination) whenever I go for microscopy. The cells are attached to seed...

28 March 2023 3,084 4 View

What is the best way to preserve plant samples intended for RNA isolation?

Hi everyone, I work with Arabidopsis thaliana. I want to know what is the best strategy to strore plant saplings after the treatment to preserve the quality of RNA (at least for a week). 1. Is it...

04 February 2023 5,443 4 View

Why am I getting an amplification in NTC (No template control) during RT-PCR?

Hello everyone! My problem is that unfortunately I am getting a good Ct value in NTC (NFW). I have tried three different primer sets (Target of three different genes) with NFW from different...

03 November 2022 7,491 9 View

How do you do methotrexate selection of Arabidopsis transformants?

So I have always used Hygromycin/Kanamycin selection for Arabidopsis but this is my first time I am using methotrexate for selection of T0 transformed plants. Plants with hygromycin resistance I...

13 December 2021 8,121 1 View

Is there an easy way to determine number of T-DNA insertion in X-generation plants of Arabidopsis?

Hi everyone! I have a few lines of a particular gene of which the earlier (T2 ratio) are not known. I want to do antibiotic selection of these seeds and I want to know which plants of next...

23 May 2021 3,912 3 View

What should be the average size of amplicon if I am using LBb1.3 (left T-DNA border) and RP (gene specific Reverse primer) against genomic DNA ?

Hello everyone! I am thinking to amplify T-DNA insertion region using LBb1.3 (left T-DNA border) and RP (gene specific Reverse Primer) against the genomic DNA of one of the T-DNA insertion mutant...

12 March 2020 1,370 3 View

Do I need to go for Dpn1 treatment?

I performed a PCR mediated deletion of my vector which was carrying an unwanted gene (Chey 183 bp). I designed two primers with desired restriction sites in order to clone any gene in place of...

09 August 2016 4,066 4 View

Why I'm getting extra protein bands with my specific protein band during Ni-NTA His-tagged protein purification?

I have expressed a protein of C. jejuni in E.coli M15 cells using pQE30 vector. My protein carries a N-terminal hexa his tag. I have purified my recombinant protein using Ni-NTA columns and I have...

07 August 2016 6,326 7 View

How can I grow two different cell lines in one plate and even they don't merge in each other?

I want to study in-vitro host-pathogen interaction and subsequent immune response. In following experiment I would like to have two different cell lines in one plate say epithelial cell line and...

29 December 2015 8,106 4 View

Why my transformed bacterial cells are not growing in antibiotic medium?

Hi, I've transformed my DH5 alpha E.coli cells with pDrive vector with my GOI. After that I plated them on ampicillin agar plate. I got good number of transformed cells (as I am expecting). There...

07 August 2015 3,060 10 View

What density of cells should I use for plating in a 35 mm cell culture dish MSC differentiation into neuronal cells?

I have been trying bFGF and Forskolin protocol to differentiate Mesenchymal Stem Cells into neuronal lineage cells using a 35 mm dish but I found my cells get overcrowded within the duration of...

08 December 2014 6,659 6 View

Is there any technique I can use to count the number of mitochondria in a cell or cell population?

I'm thinking to start a project where I have to check the number of mitochondria present in a cell population after certain physiological condition.

08 December 2014 614 3 View