CRISPR-based Cabbage CENH3 Editing

Single-guide RNAs (sgRNAs) targeting the cabbage CENH3 gene were designed using CRISPR RGEN tools (Park et al., 2015 (link)). To clone sgRNAs, primers (Supplementary Table 1) were annealed and ligated into pHSN401 (Addgene #50588) using T4 DNA ligase (NEB) and BsaI-HF (NEB) in a Golden Gate reaction. To confirm correct vector construction, we used Sanger sequencing. All vector construction procedures were performed according to Xing et al. (2014) (link).

Free full text: Click here

Stajič E, & Kunej U. (2023). Optimization of cabbage (Brassica oleracea var. capitata L.) protoplast transformation for genome editing using CRISPR/Cas9. Frontiers in Plant Science, 14, 1245433.

Publication 2023

pHSN401 T4 DNA ligase BsaI-HF

Cabbage Clone Crispr Gene Primers Single guide rnas T4 dna ligase Vector

Corresponding Organization : University of Ljubljana

Top 5 similar protocols

Variable analysis

independent variables

SgRNAs targeting the cabbage CENH3 gene

dependent variables

Correct vector construction

control variables

Primer sequences (Supplementary Table 1)
Cloning procedures (pHSN401 vector, T4 DNA ligase, BsaI-HF, Golden Gate reaction)

controls

Sanger sequencing to confirm correct vector construction

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!