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LbCas12a£¨ÓÖÃûCpf1£©ºËËáøȪԴÓÚLachnospiraceae bacterium ND2006 ¾úÖê¡£¡£¡£ ¡£¡£¡£LbCas12aÊôÓÚ2ÀàVÐÍCRISPRЧӦÂѰף¬£¬£¬£¬£¬£¬ÊÇÒ»ÖÖÒÀÀµcrRNA½éµ¼µÄºËËáÄÚÇÐø£¬£¬£¬£¬£¬£¬ÔڰбêË«Á´DNA±£´æPAM£¨TTTV£©ÐòÁеÄÇéÐÎÏ£¬£¬£¬£¬£¬£¬ÓÉcrRNA½éµ¼ÌØÒìÐÔʶ±ð²¢ÇиîË«Á´DNA£¬£¬£¬£¬£¬£¬¶øLbCas12a ÌØÒìÐÔÇиî°Ð±êµ¥Á´DNA²»ÒÀÀµPAMÐòÁС£¡£¡£ ¡£¡£¡£Í¬Ê±£¬£¬£¬£¬£¬£¬LbCas12a-crRNA¸´ºÏÎïÓ뻥²¹µ¥Á´DNA»òÕßË«Á´DNAÁ¬Ïµºó£¬£¬£¬£¬£¬£¬Æä·ÇÌØÒìÐÔssDNA·´Ê½Çиî»îÐÔ±»¼¤»î£¬£¬£¬£¬£¬£¬Í¨¹ýÉè¼ÆÁ½Í·±ê¼ÇÓ«¹â»ùÍÅ»òÆäËüС·Ö×Ó±ê¼ÇµÄµ¥Á´Reporter DNA£¬£¬£¬£¬£¬£¬¿ÉʵÏÖCas12a¶ÔDNAÄ£°åµÄ¼ì²âºÍÐźŷŴ󡣡£¡£ ¡£¡£¡£¿£¿£¿£¿£¿£¿Éͨ¹ýÓ«¹âÒǺÍÊÔÖ½ÌõÊÓ²ìЧ¹û¡£¡£¡£ ¡£¡£¡£

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LbCas12a Nuclease 1 μM*100 μL£¨100 pmol£© 1 μM*500 μL*2¹Ü£¨1000 pmol£© 1 μM*500 μL*4¹Ü£¨2000 pmol£©
LbCas12a Cleavage Buffer£¨10×£©  500 μL*1¹Ü 1mL*4¹Ü 1mL*7¹Ü

 

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10× Cleavage Buffer 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

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Fig 1. Results of Cas12a collateral cleavage at different company product.

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