Diethylpyrocarbonate

EnGen®Lba Cas12a (Cpf1, 100 μM, 20 μM) and CutSmart buffer [50 mM potassium acetate, 20 mM Tris-acetate, 10 mM magnesium acetate, 100 μg/ml bovine serum albumin (BSA), pH 7.9)] were ordered from New England Biolabs and MAGIGEN. The TranscriptAid T7 High Yield Transcription Kit and the RNeasy MinElute Cleanup Kit were obtained from ThermoFisher Scientific and Qiagen, respectively. Deoxynucleotide triphosphates (dNTPs), SYBR Green, diethylpyrocarbonate (DEPC)-treated water, eight strip real-time PCR tubes and caps, magnesium chloride, reporter, DNA sequences, uracil-DNA glycosylase (UDG) and PCR mix were purchased from Sangon Biotech Inc. (Shanghai, China) and Tsingke Biotechnology Co., Ltd (Beijing, China). The crRNAs used were synthesized with the TranscriptAid T7 High Yield Transcription Kit or obtained from Beijing Genomics Institution (Beijing, China). The nucleic acid extraction and purification kit was purchased from Burning Rock Biotech Ltd. (Guangzhou, China). N,N,N',N'-Tetramethylethylenediamine (TEMED) and 30% acrylamide/bis solution were provided by Sigma-Aldrich (St. Louis, MO, USA). DNA loading buffer (6×) and Gel Red nucleic acid dye were ordered from TaKaRa Biotech (Dalian, China). All chemical reagents were of analytical grade, and RNase-free water was used throughout this study.