Crispr nickase, how to design the homology arm?

14 December 2020 0 2K Report

Dear All,

I am new in the field of the crispr. As far as I know, nickase crispr need two sgRNA. However, I dont know how to design the homology arm in the nickase crispr case. Can u guys give me an example, so I can finish the tasks?

Badges
Science topic

Similar topics
Psychology

More Ting Chung Chau's questions See All

Regarding LBM in Boiling?

What should be the equation for halfway bounce back boundary condition for flow field with forcing term?

08 August 2024 7,938 1 View

In boiling using pseudopotential LBM density and temperature used in P-R EOS should be change with each iteration or taken as saturated value?

Peng Robinsion EOS,P=[ρRT/(1-bρ)]-[(aα(T)ρ2)/(1+2bρ-b2 ρ2 )] 〖α(T)=[1+(0.37464+1.54226ω-0.26992ω^2)(1-√(T⁄(Tc)))]〗^2, where ω=0.344 for water, a= 2/49, b=3/21 & R=1. ψ(ρ)=√(2(P-ρcs2 )/(c0...

01 April 2024 5,631 1 View

Which statistical analysis method should I use for 2 IVs, 1 mediator and 1 DV?

I am currently working on a research with 2 IVs (both are categorical with 2 levels), 1 mediator and 1 DV. Apart from the mediation effects, I am also interested in the interaction effect of the...

24 March 2024 566 0 View

What are the reasons for the LCMS autosampler broken (bending needle, sample transport broken)?

Hello everyone! I would like to ask you about the reasons for autosampler break. The previous day, I used the LCMS machine and I did not put a sample in the rack at which the autosampler came...

30 January 2024 2,487 2 View

Silver chloride photocatalysis?

After AgCl photocatalytic degradation of Rhodamine, it turns black and has poor stability. How to make it more stable under light irradiation

15 January 2024 9,357 2 View

SAP2000 V20 Pushover cannot converge, how to resolve?

I have no issues with my plastic hinge settings, and it converges well in V14. However, when opening the file in V20, it fails to converge. How should I address this? I have set the hinge...

19 December 2023 4,911 4 View

Immobilization of photocatalysts?

What I am doing recently is to load bismuth oxybromide on foam nickel. The hydrothermal method is adopted. After hydrothermal treatment, foam nickel is dried in an 80 degree oven to degrade...

25 October 2023 3,377 1 View

Why do we get E0-0 energy value by the crossing point of normalized absorption spectra and emission spectra using 1240/λ ?

why do we get E0-0 energy value by the crossing point of normalized absorption spectra and emission spectra using 1240/λ ?

02 October 2023 6,971 1 View

why did all the lung cancer cells transfected with px459 plasmid die after two-day antibiotic screening?

I have applied Lipofectamine 3000 and Versatile DNA/siRNA transfection reagent jetPRIME® to transfect PX459 plasmid into A549 and NCI-H1975, all the operations are performed according to the...

01 September 2023 3,745 1 View

Interpreting kinetic values in IntEnzyDB for multi-substrate reactions?

Hello! I'm currently using the IntEnzyDB (https://intenzydb.accre.vanderbilt.edu/kinetics/list) for a machine learning project. During this process, I noticed that for some observations in the...

01 August 2023 6,945 3 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

Question about ELISA with the "Cattle IGF 1 (Insulin like growth factor) ELISA Kit" Cat: ELK5630?

Dear fellow researchers, We are conducting an ELISA with the "Cattle IGF 1 (Insulin-like growth factor) ELISA Kit" (Cat: ELK5630). Samples and reagents were prepared following the instruction...

13 June 2024 9,473 0 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