Empty Crispr/Cas9 donor plasmid?

More Mirza Muhammad Fahd Qadir's questions See All

A good fluorophore conjugated P2Y1 receptor antibody out there?

Has anyone used a good P2Y1 receptor antibody conjugated to 488 or 647? For FACS and cell sorting!! Thanks!!!

07 August 2016 7,698 0 View

What is a good set of antibodies for mesenchymal/fibroblast cells and endothelial cells. Considering I want to eliminate these populations in FACS?

Hello! I want to sort out mesenchymal and endothelial populations (essentially the junk) from a population of cells in live Acinar Pancreatic tissue. Can anyone give me suggestions on what array...

05 June 2016 8,429 1 View

Why am I getting overexposure on my immunoblotting?

Hi! Im performing a westren blot, but my immunoblotting isnt going well. The proteins transfer perfectly on the PVDF membrane, but when I incubate the membrane with my primary abs, the signal...

06 July 2014 2,273 13 View

Is it possible to make a cell lysis buffer without PMSF?

Can someone please tell me how to make a cell lysis buffer without PMSF I have sodium deoxycholate but no PMSF. I also have NP-40. Please tell me an appropriate recipe one which does not include...

06 July 2014 1,717 7 View

In animal models, particularly BALB/c mice, how can I induce arthritic induction using collagen type-II?

Arthritic induction is required by the protein rich in joints.

05 June 2014 4,976 5 View

Hi! An excellent project. I would love to connect with you and discuss our research as well. I feel we may have similar interests. Can we?

Our lab studies the effect of BMP7 on pancreatic-progenitor like cells, however we have not studied the effect of this on bone mineralization. Maybe we could collaborate in this regard!

01 January 1970 7,008 2 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View