What are the hairpin free energy limits to design crispr sgRNA?

More Ruud Grootens's questions See All

Has anyone used a Phalloidin stain that permeates the cell membrane of living cells (rat cardiomyocytes)?

In this particular experiment, it is not possible to perform any permeabilization protocol that weakens the sarcolemma or cytoskeleton, so I would have to rely on finding a Phalloidin that can...

09 January 2018 2,847 4 View

Do the workers that apply, but don't get the job get enough focus?

Is there any articles or tips? I'm doing a research on internal recruitment. What happens to the workers that don't get the job. And don't get the feedback they need. Motivation? Turnover? I need...

26 January 2015 9,969 2 View

Do you have recent literature about prevalence of trauma and PTSD among general mental health care populations?

I am looking for recent figures about prevalence of trauma and PTSD in the different levels of populations, but especially among general mental health populations, like out-patients clinics. Does...

19 November 2014 4,036 10 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

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 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

What is the most effective method for knocking out a gene in Candida albicans?

I have attempted various methods, including homozygous gene disruption and CRISPR-Cas9, but have not achieved the desired results. It appears that the gene of interest is situated in a...

15 July 2024 3,977 1 View

What would you do next?

Studying a specific monogenetic disease based on mutations in different individual genes. I try to recreate the disease phenotypes of donors in vitro. I knocked out a gene x with above 90%...

12 July 2024 6,313 0 View

Are there any information about protocols of CRISPR in leishmania strains?

I am looking for a CRISPR protocol for application in leishmania strains

08 July 2024 9,732 1 View

Is it possible to generate a (semi-)permanent "transcription bubble" within a replicable plasmid?

I was wondering if it is possible to form a permanent open "ssDNA bubble" similar to a transcription bubble (>13 nucleotides) within E. coli. These criteria are important: 1. Open ssDNA...

07 July 2024 8,275 5 View

There is no colonies after Ligation reaction and transformation ?

I am currently investigating the CRISPR 9 case to target my gene of interest for knockout. The vector being used is lentiCRISPR v2puro. I digested and dephosphorylated 5ug of the lentiviral...

03 June 2024 2,877 7 View

How can I isolate the desired cell population after making a point mutation by using CRISPR/Cas 9? Are there any ways to increase the rate of HDR?

Now, I am using CRISPR/Cas 9 system to make a point mutation at THBD gene. I am wondering how can I isolate the desired cell population after making a point mutation by using CRISPR/Cas 9? Are...

30 May 2024 9,993 0 View