pFH6_new
(Plasmid #105866)

• Purpose: more efficient version of pFH6; Subcloning of any sgRNA via BbsI site
• Depositing Lab: Andreas Weber
• Publication: Hahn et al Bio Protoc. 2017 Jul 5;7(13):e2384. doi: 10.21769/BioProtoc.2384. eCollection 2017 Jul 5.

Backbone

• Vector backbone: pGEM T-easy
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 3017
• Total vector size (bp): 3620
• Vector type: Plant Expression, CRISPR, Synthetic Biology ; Subcloning

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: U6-26p::sgRNA scaffold
• gRNA/shRNA sequence: can be inserted via BbsI sites
• Species: A. thaliana (mustard weed), Synthetic

Resource Information

• Supplemental Documents:
  • pFH6 new genebank.gb
  • Cloning Protocol for pFH6_new and pUB-Cas9
• A portion of this plasmid was derived from a plasmid made by: Peter Hegemann (HU Berlin)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The new version of pFH6 (pFH6_new) provides > 10 times higher mutation efficiencies than the original version. In combination with pUB-Cas9, homozygous knockout plants can be created in Arabidopsis in the T2 generation at efficiencies of 50-100%.

For cloning procedure, please use the attached protocol which is derived from Hahn et al., 2017 ( http://www.bio-protocol.org/e2384 ).