Anindhya Sundar Das

9 Questions 13 Answers 0 Followers

Questions related from Anindhya Sundar Das

How metaphor agarose can be used to genotype Indels introduced by single gRNA?

Dear all, We have introduced double stranded break in our gene (exonic) using a single gRNA. We have also checked efficiency of the cut by T7E1 assay. Now we are trying to genotype invidual clones...

06 June 2018 5,411 3 View

How can I tag mRNA by cloning inside a cell?

Hi I am interested to check a particular mRNA and a protein interaction/ localization inside cells. For this reason, I want to tag the transcript. Beside RNA-FISH or any such hybridization...

11 November 2017 8,742 0 View

In there any report on Lipofectamine RNAiMAX mediated transfection of CRISPR-Cas9 plasmid in THP-1 cells?

Hi..is there any literature on successful gene knock out in THP-1 monocytes via CRISPR-Cas9 using Lipofectamine RNAiMAX? Thanks in Advance

03 March 2017 4,752 1 View

Any suitable protocol for Ni-NTA based pull down and interaction study?

Hi..I have over-expressed a protein (tagged with His) via retroviral transduction method in THP-1 cells. Now I am interested to check if there is any binding partner with my tagged protein in a...

12 December 2016 2,712 4 View

How can I separate ribosome-free cytosol using sucrose cushion?

I intend to separate ribosomal and non-ribosomal fractions of cytosol using 20% sucrose cushion. I have already run the cell lysate on top of the 20% sucrose solution for 2 hrs at 1,39,000 g. I...

06 June 2016 1,163 2 View

How can I search the expression of particular set of genes in NCBI GEO database?

I am interested in searching the gene expression profile of a particular set of genes in a specific disease condition like atherosclerosis from NCBI GEO database. As I have almost no background in...

07 July 2015 662 7 View

Why CD11b and CD14 mRNA expression is getting down after 6hrs of rest?

I have converted THP1 monocytes to macrophage using 5 ng/ml PMA for 48 hrs in complete RPMI1640.PMA Containing media was discarded and cells were washed with sterile DPBS for 3 times and given...

04 April 2015 5,151 2 View

Why are my THP1 cells forming clumps?

I am culturing THP1 cells in RPMI 1640 (as advised by ATCC formulation, except 2ME). Unfortunately my cells are forming clumps a few hours after seeding (at 1* 10^5 cells/ml) before even going to...

03 March 2015 214 10 View

Why are my PMA differentiated THP1 cells not responding to LPS induction?

I have differentiated THP1 monocytes into macrophage using 5 ng/ml PMA for 48 hrs. Then after washing twice with DPBS i have kept the diffrerentiated cells for 3 hrs in serum free media. Then I...

03 March 2015 1,359 8 View