DNA transfection Mammalian cells

Get tips on using pAC2-dual-dCas9VP48-sgExpression to perform CRISPR Human - Activation IL1RN

Get tips on using pAC2-dual-dCas9VP48-sgExpression to perform CRISPR Human - Activation SOX2

Get tips on using pAC154-dual-dCas9VP160-sgExpression to perform CRISPR Human - Activation HIV-1 5′ LTR

Get tips on using MISSION® shRNA SOX2 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans SOX2 lentiviral particles

Get tips on using MISSION® shRNA SOX6 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans SOX6 lentiviral particles

Get tips on using MISSION® shRNA ZEB2 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans ZEB2 lentiviral particles

Get tips on using MISSION® shRNA ZEB1 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans ZEB1 lentiviral particles

Get tips on using HTRA2 MISSION shRNA Lentiviral Transduction Particles HtrA serine peptidase 2 to perform shRNA gene silencing Mouse - FL83B HtrA2

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.