How many bands do I expect to observe after agarose gel electrophoresis of crRNA:tracrRNA duplex?

Why is my ASO degraded quickly on agarose gel?

I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...

06 August 2024 3,130 2 View

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...

01 August 2024 4,673 4 View

MAGeCK Plotting Error: Logarithmic Axis Must Have Positive Limits?

I have been running a MAGeCK test command on the terminal for a CRISPR screen to rank sgRNAs and genes based on the read count tables. However, I get only the plot of the top-ranked genes...

28 July 2024 9,811 1 View

Why knockouts for same genes from two different gRNA are showing different result?

Why knockouts for same genes from two different gRNA are showing different result? I followed CRISPR-CAS9 gene editing technology.

25 July 2024 8,313 1 View

Why my gel electrophoresis has smearing?

Greetings. I’m currently running a nested pcr for giardia. My mastermix comprised of 3mM Mgcl2, 5unit of taq polymerase, 0.2uM of forward and reverse primers each respectively and 0.2mM of dNtp...

22 July 2024 9,761 5 View

Is possible to edit a gene present in a plasmid by Crispr Cas9?

Hello everyone! Someone working with Crispr Cas9. I have a question. Is it possible to edit a gene that is present on a plasmid (in this case with a low copy number)? In this case, is it possible...

22 July 2024 7,073 2 View

How should we increase the quality of RNA extraction?

I’m having difficulty achieving high RNA integrity in my samples. Although the 260/280 and 260/230 ratios are satisfactory after RNA extraction, the RNA samples show signs of degradation when...

22 July 2024 155 4 View

Why my gel electrophoresis have shadow bands? Please see the attached picture for the gel electrophoresis ?

Sometimes I see the shadow like bands and its not true band. I want to know that what's the reason for it. I am using 2% gel for running genotyping samples I have uploaded the gel picture in both...

19 July 2024 148 6 View

Less substrate signals towards the last lanes of the Native PAGE?

I have been running native page for FAM DNA substrate ( fluorescence samples) for protein DNA binding reaction. Binding is there but towards the end of the lane , I am loosing signals...

17 July 2024 6,213 4 View

How to activate 4E8 T cells with plate-bound anti-CD3 for a genome-wide CRISPR screen?

It was assumed that 4E8 T cells should be isolated and activated using plate-bound anti-human CD3, after which would be used for a genome-wide CRISPR screen. (Ref: Genome-wide CRISPR Screens in...

17 July 2024 3,371 0 View