Gene editing using Cas9 RNP?

Hi everyone, Im trying targeted gene editing in citrus protoplast through PEG-mediated transformation. Im using sgRNA+Cas9 RNP complex for DNA free genome editing. But im not successful in...

06 July 2019 1,357 1 View

How to design and insert my target sequence into sgRNA scaffold?

Dear All, I want to design sgRNA target sequence and insert into scaffold portion already present in pRGEB31 binary vector. Any experimental protocol for that? I want to knock out a disease...

05 June 2018 4,787 2 View

Anyone knows in vitro digestion of DNA with Cas9?

I am going to check my gRNA efficiency by in vitro digestion of DNA with Cas9. I read the protocol from NEB website but have some doubts regarding that. I want to know whether i should do in vitro...

05 June 2018 9,806 0 View

Standard protocol for plant tissue preparation to visualize GFP?

10 November 2016 1,287 5 View

Is vacuum infiltration really effective for agrobacterium mediated transformation in seeds of recalcitrant plants?

08 September 2016 6,703 3 View

Antioxidant enzymes in cotton seedlings?

06 July 2016 4,830 3 View

Any suggestion regarding protein sample preparation for 2D gel electrophoresis?

05 June 2016 283 4 View

Protein expression in cotton seedlings under abiotic stress?

03 April 2016 8,114 7 View

Does bubble formation affect separation of proteins in 2D electrophoresis?

03 April 2016 448 13 View

Good protocol for silver staining?

02 March 2016 1,862 6 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 3 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View