Protein expression and purification Bacteria Escherichia coli

Get tips on using pANC210 (xylB) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylB

Get tips on using pANC209 (xylA) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylA

Get tips on using pFastBac1-TEV-TRPV1 to perform Protein Expression Eukaryotic cells - HEK293 TRPV1

Get tips on using pCMV-HA2/DKK1 to perform Protein Expression Eukaryotic cells - HEK293 DKK1

Get tips on using pgMAX system-DsRed2 to perform Protein Expression Eukaryotic cells - HEK293 DsRed2

Get tips on using pCEP-4-MBP to perform Protein Expression Eukaryotic cells - HEK293 BM40

Get tips on using pMIR-REPORT™ miRNA Expression Reporter Vector System to perform Reporter gene assay luciferase - HEK 293 human embryonic kidney cells

Get tips on using Qproteome Nuclear Protein Kit to perform Protein enrichment Soluble nuclear proteins

Bacterial culture is a process of letting bacteria multiply in a controlled fashion (temperature, humidity, oxygen content or shaking), in a predetermined culture medium (antibiotic resistance to obtain homogenous clones). It is an important step, especially during cloning, as a single cell can be grown homogeneously (on semi-solid or in liquid conditions) to obtain colonies. As mentioned, bacteria can be cultured in broth cultures (Luria broth or LB) or Petri dishes (Agar plates). A specific antibiotic can be added to the broth or agar plates in order to grow bacteria which have the gene insert conferring its resistance to that antibiotic. Following points are necessary to consider for optimal growth conditions: 1. In general, most bacteria grow well at 37C, but there are some strains which require growth temperatures between 25-30C. 2. It is ideal in broth cultures to fill the flask to ⅓ or less of the total flask volume for optimal aerobic growth. 3. Shaking speeds between 140-180 rpm are appropriate to ensure aeration and that the cells are surrounded by fresh media, and do not settle.

Bacterial culture is a process of letting bacteria multiply in a controlled fashion (temperature, humidity, oxygen content or shaking), in a predetermined culture medium (antibiotic resistance to obtain homogenous clones). It is an important step, especially during cloning, as a single cell can be grown homogeneously (on semi-solid or in liquid conditions) to obtain colonies. As mentioned, bacteria can be cultured in broth cultures (Luria broth or LB) or Petri dishes (Agar plates). A specific antibiotic can be added to the broth or agar plates in order to grow bacteria which have the gene insert conferring its resistance to that antibiotic. Following points are necessary to consider for optimal growth conditions: 1. In general, most bacteria grow well at 37C, but there are some strains which require growth temperatures between 25-30C. 2. It is ideal in broth cultures to fill the flask to ⅓ or less of the total flask volume for optimal aerobic growth. 3. Shaking speeds between 140-180 rpm are appropriate to ensure aeration and that the cells are surrounded by fresh media, and do not settle.