Empty Crispr/Cas9 donor plasmid?

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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 7,804 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 8,517 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 2,403 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 1,844 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 5,065 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 7,094 2 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

What is the acceptable p-value cutoff for GO enrichment analysis ?

I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...

31 July 2024 225 2 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

Issues with Protein Expression in Transfected cells?

Hello I am trying to create a stable cell line in HEK293 via Lipofectamine 3000 transfection. My plasmid is a CD63-IL10-GFP construct with Puromycin resistance. I am successful with the...

30 July 2024 6,648 1 View

Inquiry on Maximum Nucleic Acid Volume for 2.5 mL Liposome Solution?

I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...

30 July 2024 6,420 1 View

Does anyone have experience electroporating SKOV3 cells with a large plasmid using Neon transfection?

I have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?

29 July 2024 3,229 1 View