Author: Long T. Nguyen; Brianna M Smith; Piyush K. Jain
Title: Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection Document date: 2020_4_14
ID: n5kpvsvg_34
Snippet: Nonspecific cleavage activity of Cpf1 was activated by incubating Cpf1, crRNA, and DNA activator with a concentration of 100 nM:100 nM: 2nM in 1X NEBuffer 2.1 buffer at 37 o C for 30 minutes. M13mp18 was then added to the 30 ïL reaction mixture and incubated for an additional 45 minutes. A fraction of the reaction was taken out every 5 minutes, quenched in 6X purple gel loading dye (New England Biolabs Inc.), and subsequently analyzed in 1% aga.....
Document: Nonspecific cleavage activity of Cpf1 was activated by incubating Cpf1, crRNA, and DNA activator with a concentration of 100 nM:100 nM: 2nM in 1X NEBuffer 2.1 buffer at 37 o C for 30 minutes. M13mp18 was then added to the 30 ïL reaction mixture and incubated for an additional 45 minutes. A fraction of the reaction was taken out every 5 minutes, quenched in 6X purple gel loading dye (New England Biolabs Inc.), and subsequently analyzed in 1% agarose gel (Fisher Scientific) 1 .
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