(3-Chloro-4-fluorophenyl)-7-me

Assisted large fragment insertion by Red/ET-recombination (ALFIRE)―an alternative and enhanced method for large fragment recombineering

containing chloramphenicol (Cm, 20 μg/ml)/streptomycin (Stp, 75 μg/ml) and purified using the Large construct kit (Qiagen, Valencia, CA, USA). PCR PCR reactions using oligonucleotides carrying HAs were performed using

Purification of GST fusion proteins in E.coli GST融合蛋白纯化（方法四）筛选表达

proteins).-Note: Crude sonicates can be screened for the relative level of expression of GSTfugion proteins using the GST substrates CDNB (1-chloro-2,4-dinitrobenzene).Centrifuge in a microcentrifuge for 5 min to remove insoluble materials.Save a10 μl

GST融合蛋白纯化--筛选表达株

using the GST substrates CDNB (1-chloro-2,4-dinitrobenzene).Centrifuge in a microcentrifuge for 5 min to remove insoluble materials.Save a10 μl aliquots of the insoluble material for analysis by SDS-PAGE.Transfer the supernatants to fresh tubes,Add 20 μl