LbCas12a 是一种crRNA介导的核酸酶，在PAM（Protospacer Adjacent Motif）序列（TTTV）存在的情况下，能够识别并切割双链DNA（dsDNA），结果可以顺利获得荧光计和试纸观察。
* 仅供科研使用，不适用于人体或动物，包括临床、治疗或诊断用途。
产品基本信息
LbCas12a（又名Cpf1）核酸酶来源于Lachnospiraceae bacterium ND2006 菌株。LbCas12a属于2类V型CRISPR效应蛋白，是一种依赖crRNA介导的核酸内切酶，在靶标双链DNA存在PAM（TTTV）序列的情况下，由crRNA介导特异性识别并切割双链DNA，而LbCas12a 特异性切割靶标单链DNA不依赖PAM序列。同时，LbCas12a-crRNA复合物与互补单链DNA或者双链DNA结合后，其非特异性ssDNA反式切割活性被激活，顺利获得设计两端标记荧光基团或其它小分子标记的单链Reporter DNA，可实现Cas12a对DNA模板的检测和信号放大。可顺利获得荧光仪和试纸条观察结果。
产品组分
| 组分 | EDE0005-100 | EDE0005-1000 | EDE0005-2000 |
| LbCas12a Nuclease | 1 μM*100 μL（100 pmol） | 1 μM*500 μL*2管（1000 pmol） | 1 μM*500 μL*4管（2000 pmol） |
| LbCas12a Cleavage Buffer（10×） | 500 μL*1管 | 1mL*4管 | 1mL*7管 |
反应体系
| 组分 | 终浓度 | 体积 ( μL) |
| 10× Cleavage Buffer | 1× | 3 |
| 1 μM LbCas12a Nuclease | 33 nM | 1 |
| 1 μM crRNA | 33 nM | 1 |
| 4 μM ssDNA Reporter | 400 nM | 3 |
| 100 nM DNA target | 3.3 nM | 1 |
| DEPC H2O | ||
| Total | 30 μL | 30 μL |
反应条件
使用实时荧光定量PCR仪或恒温扩增仪检测荧光信号，37℃反应，每30 sec采集一次荧光信号。
竞争性能交叉分析
横坐标为反应时间，纵坐标为 Bio-rad CFX96荧光PCR仪检测结果。由图可见，相同的条件下，可见本品切割效率显著高于同类产品。
Fig 1. Results of Cas12a collateral cleavage at different company product.
储存条件及有效期
有效期1年，保存条件-80°C。为避免反复冻融，Cas酶开封使用过后请置于-20°C；建议根据使用次数进行分装使用，避免温度多次变化导致酶活性降低。
* 研究用途免责声明：本内容基于公开的研究数据、生物信息学资源及计算分析生成，仅供研究参考。
已验证文献
Direct microRNA detection via topologically engineered CRISPR/Cas12a cascade amplification assay
IF=4.6
Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy
Establishment of One-Pot ERA-CRISPR/Cas12a-Based Rapid Visual Assays and a TaqMan Quantitative PCR Assay for Lactococcus garvieae
Microorganisms
Asy-RPA/PCR combined with One-crRNA-CRISPR/Cas12a for simultaneous detection of multiple Clarithromycin resistance mutations in Helicobacter pylori
IF=4.6
Nanomedicine: Nanotechnology, Biology and Medicine
A ratiometric and trans-cleavage-triggered feedback CRISPR/Cas12a cascade for amplification-free microRNA detection using a multifunctional switch
Microchemical Journal
Visual CRISPR/Cas12a-mediated Rapid Detection of Gyrovirus galga1
IF=5.4
Pakistan Veterinary Journal
DNAzyme-driven SDA reaction regulates CRISPR/Cas12a for highly sensitive and selective analysis of underexpressed miRNA
IF=6
Analytica Chimica Acta
Field-parallel six-sample microfluidic detection of plant viruses via raffinose-assisted one-pot LAMP-CRISPR/Cas12b
IF=13
Journal of Advanced Research
cliCRISPR: crRNA-limited CRISPR/Cas12a system for multiplexed detection
IF=10.5
Biosensors and Bioelectronics
A multifunctional switch for label-free CRISPR/Cas12a sensor with self-driven amplification
IF=4.4
Synthetic and Systems Biotechnology
A dual fluorescence channel RAA-based CRISPR-Cas12a/Cas13a system for highly sensitive detection of Gyrovirus galga1 and Gyrovirus homsa1
IF=5.4
Taylor & Francis
CRISPR/Cas12a-mediated DNAzyme/split-aptamer cascade for label-free detection of site-specific DNA methylation
IF=10.5
Biosensors and Bioelectronics
A small library crRNA-enhanced CRISPR-Cas12a system for ultrasensitive point-of-care test of hantavirus M gene
IF=4.9
Sensing and Bio-Sensing Research
Point-of-care testing diagnosis of African swine fever virus by targeting multiple genes with enzymatic recombinase amplification and CRISPR/Cas12a System
IF=4.8
Frontiers in Cellular and Infection Microbiology
Rapid and multiple visual detection of Fasciola hepatica in feces via recombinase polymerase amplification integrated with CRISPR/Cas12a technology
IF=8.5
International Journal of Biological Macromolecules
A super convenient and specific CRISPR/Cas12a diagnostic platform for toxigenic Burkholderia gladioli based on virulence genes
IF=5.6
Food Control
Dumbbell probe initiated multi-rolling circle amplification assisted CRISPR/Cas12a for highly sensitive detection of clinical microRNA
IF=10.7
Biosensors and Bioelectronics
Optimization of CRISPR/Cas12a detection assay and its application in the detection of Echinococcus granulosus
IF=2
Veterinary Parasitology
PAIT effect: Padlock activator inhibits the trans-cleavage activity of CRISPR/Cas12a
IF=10.7
Biosensors and Bioelectronics
Rapid and Simple Detection of Burkholderia gladioli in Food Matrices Using RPA-CRISPR/Cas12a Method
IF=5.1
Foods
A Rapid RPA-CRISPR/Cas12a Detection Method for Adulteration of Goat Milk Powder
IF=5.1
Foods
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