Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos

. 2021 Dec 23;3(1):101058. doi: 10.1016/j.xpro.2021.101058

Here we review the material specifically needed for RfxCas13d protein purification and E3 buffer:

Solution	Composition	Storage
Liquid LB	2% (w/v) LB in ddH₂O (autoclaved)	25°C, 1 month
LB-Agar-Chl plates	2% (w/v) agar, 15 μg/mL Chloramphenicol in liquid LB (autoclaved)	4°C, 1 month
LB-Agar-Chl-Kan plates	2% (w/v) agar, 15 μg/mL Chloramphenicol, 25 μg/mL Kanamycin A in liquid LB (autoclaved)	4°C, 1 month
TSS Buffer	See below	4°C, 12 months
15 mg/mL Chloramphenicol	1.5% (w/v) Chloramphenicol in ethanol	−20°C, 6 months
25 mg/mL Kanamycin A	2.5% (w/v) kanamycin sulfate in ddH₂O	−20°C, 6 months
1 M IPTG	23.83% (w/v) IPTG in ddH₂O	−20°C, 6 months
1 M DTT	15.4% (w/v) DTT in ddH₂O	−20°C, 1 month
Tris·HCl 20 mM pH 7.5	0.24% (w/v) Tris in ddH₂O, neutralized with HCl	25°C, 6 months
HEPES·KOH 1M pH 7.5	23.83% (w/v) HEPES in ddH₂O, neutralized with KOH	25°C, 6 months
Lysis/Binding Buffer	See below	25°C, 6 months
Elution Buffer	See below	25°C, 6 months
Dialysis Buffer	See below	25°C, 6 months
E3 buffer	See below	25°C, 1 month