What are Stbl2 cells?
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MAX Efficiency® Stbl2™ Competent Cells are high-efficiency chemically competent cells specifically designed for cloning unstable inserts. In addition to recA1, a unique set of genetic markers allow for stable cloning of direct repeat and retroviral sequences and tandem array genes.
Why use Stbl3?
When using lentiviral vectors, Stbl3 has advantages. If your lentiviral vector is big, then Stbl3 for sure will be better to use. Stbl3 have higher transformation efficiency with larger plasmids, and also have less mutation and recombination rates.
How do you make a competent cell Stbl3?
Thaw, on ice, one vial of One Shot™ Stbl3™ chemically competent cells for each transformation. 2. Add 1 to 5 μL of the DNA (10 pg to 100 ng) into a vial of One Shot™ cells and mix gently. Do not mix by pipetting up and down.
How do Stbl3 cells grow?
Preparation: Use glycerol stock or stock of existing competent Stbl2 to streak cells for single colonies on LB plate without antibiotics and grow over night in bacterial incubator (37°C). Store plate at 4°C. Plate should be good for at least 2 weeks.
How do you make a competent bacterial cell?
Hence, in order to make bacteria capable of internalizing the genetic material, they must be made competent to take up the DNA. This can be achieved by making small holes in bacterial cells by suspending them in a solution containing a high concentration of calcium.
How do you make a cell more competent?
Cells must remain cold for the rest of the procedure: Transport tubes on ice and resuspend on ice in the cold room. Decant supernatant and resuspend the cells in 1/4 original volume (87.5 ml) ice cold 100 mM MgCl2. Hold on ice for 5 minutes. Transfer the cells to pre-chilled sterile large centrifuge bottles.
How do you calculate transformation efficiency of competent cells?
Transformation efficiency is the efficiency by which cells can take up extracellular DNA and express genes encoded by it. This is based on the competence of the cells. It can be calculated by dividing the number of successful transformants by the amount of DNA used during a transformation procedure.
Why do we need to use BL21 DE3 and not BL21 cells for expression with IPTG?
IPTG is required to maximally induce expression of the T7 RNA polymerase in order to express recombinant genes cloned downstream of a T7 promoter. BL21(DE3) is suitable for expression from a T7 or T7-lac promoter or promoters recognized by the E.
What are max efficiency® stbl2™ competent cells?
MAX Efficiency® Stbl2™ Competent Cells are high-efficiency chemically competent cells specifically designed for cloning unstable inserts. In addition to rec A1, a unique set of genetic markers allow for stable cloning of direct repeat and retroviral sequences and tandem array genes. MAX Efficiency® Stbl2™ Competent Cells offer:
What are stbl4™ competent cells?
ElectroMAX™ Stbl4™ Competent Cells are specifically designed for cloning unstable inserts. As electrocompetent cells, they have one of the highest transformation efficiencies available (>5×10 9 cfu/µg), making them ideal for generating cDNA and genomic libraries and for cloning unstable inserts. These electrocompetent E. coli:
Does stbl2 improve stability when cloning retroviral or direct repeat sequences?
Invitrogen Stbl2 and Stbl4 competent cells are both designed to improve stability when cloning retroviral or direct repeat sequences. In a series of experiments, Stbl2 was compared directly to several other strains also known for increasing stability of retroviral and tandem repeat inserts. An article in the Focus Journal (Issue 16.3, p.
What is the role of stbl2 cells in toxic gene transformation?
These cells carry the lacIq repressor that represses expression from the lac promoter and so allows cloning of toxic genes. Keep in mind that in the absence of IPTG, blue-white screening cannot be performed. – Try using Stbl2 cells for the transformation.