1、cell grown to stationary phase in 100mL of YM medium.
2、protoplast preparation.
3、Collected by centrifugation,washed twice with 50mM EDTA pH7.5 and suspended in 1mL by the 50mM EDTA .
4、This suspension was then mixed with 3mL of 1%low melting point agarose in 125mM EDTA pH7.5 which was prewarmed to 42 ℃。bs认证
5、Mixture incubated at 37℃for 30min, distributed in 100μL molds(LKB) and allowed to solidify for 30 min at 4℃。
6、Then immersed in 5mL of a solution containing enzyme(4mg Novozyme234 per mL)in LET buffer.(500mM EDTA,7.5% 2-mercaptoethanol巯基乙醇，10mM Tris pH7.5), 37℃16h.
脂肪酸酰胺7、Plugs immersed in 5mL of NDS solution(2mg/mL proteinase K in 500mM EDTA,1% lauroyl sarcosine月桂酰肌氨酸，10mM Tris-HCl,pH7.5 ) 50℃24h.
8、Repeated incubation 5ml freshly prepared NDS solution,50℃16h.蒸汽回收机 9、Washed the plugs twice with 50mM EDTA pH7.5 keep in this solution at 4℃until use.
1、cell grown to stationary phase in 100mL of YM medium.
2、protoplast preparation.
3、Collected by centrifugation,washed twice with 50mM EDTA pH7.5 and suspended in 1mL by the 50mM EDTA .
4、This suspension was then mixed with 3mL of 1%low melting point agarose in 125mM EDTA pH7.5 which was prewarmed to 42 ℃。机控网
永磁
同步电机反电动势
5、Mixture incubated at 37℃for 30min, distributed in 100μL molds(LKB) and allowed to solidify for 30 min at 4℃。
6、Then immersed in 5mL of a solution containing enzyme(4mg Novozyme234 per mL)in LET buffer.(500mM EDTA,7.5% 2-mercaptoethanol巯基乙醇，10mM Tris pH7.5), 37℃16h.
7、Plugs immersed in 5mL of NDS solution(2mg/mL proteinase K in 500mM EDTA,1% lauroyl sarcosine月桂酰肌氨酸，10mM Tris-HCl,pH7.5 ) 50℃24h.
搁物架
8、Repeated incubation 5ml freshly prepared NDS solution,50℃16h.
9、Washed the plugs twice with 50mM EDTA pH7.5 keep in this solution at 4℃until use.
豫公网安备41160202000603
站长QQ:729038198
关于我们
投诉建议