STBL3 感受态细胞(化转)/ STBL3 chemica

STBL3 chemically Competent Cells

 

STBL3 chemically Competent Cells Components

STBL3锛?nbsp;                                                  100μl/tube

pUC19 (control vector锛?0pg/μl):                    10μl

Storage at                             -80鈩冿紙6month锛?/span>

 

STBL3 chemically Competent Cells Genotype

F- mcrB mrr hsdS20(rB-, mB-) recA13 supE44 ara-14 galK2 lacY1 proA2 rpsL20(Strr) xyl-5 λ-leu mtl-1 endA1+

 

STBL3 chemically Competent Cells Description

Stbl3, derived from HB101 E. coli strain, is a strain recommended for the lentivirus vector system. The genome contains recA13 mutation of recombinant enzyme, which can effectively inhibit the recombination of long terminal repeat region, reduce the probability of false recombination, but does not contain nuclease endA1 mutation, and has high nuclease content in vivo. When we extract plasmid, we must use Plasmid Extraction Kit. This strain has streptomycin resistance and does not exist lacIqZ Delta M15. It can not be screened by Yu Lan and leukoplakia. The Stbl3 receptive cells were made by special process, and the conversion efficiency of >108cfu/ mu g DNA was detected by pUC19.

 

STBL3 chemically Competent Cells Transformation Protocol
1. Please store STBL3 chemically Competent Cells in -80°C for late use.
Take it then insert the tube in the ice immediately for 5 minutes.
2. Add your plasmid DNA (vector) to the cell mixture. Carefully flick the tube 4–5 times to mix cells and DNA. Do not vortex. Place the mixture on ice for 25 minutes, Do not mix.
3. Place it in the 42°C water for 45 seconds, then immediately transfer the tube back into ice for 2min. Do not mix.
4. Add 900ul LB or S.O.C nutrient medium (No antibiotic). then put on the Constant temperature shaker (225 rpm) for 1 hours at 37°C or (225 rpm) for 1.5 hours at 30°C ^[1].
5.Pellet the mixture by centrifugation at 5000rpm/min for 1min. Pour off about 100ul of cell supernatant, then resuspend the cell pellet in the remaining medium by gently vortexing the tube. Spread the cell on the LB or S.O.C nutrient medium Resistance plates by antibiotic, then inverted and Incubate overnight at 37°C or 30°C ^[2].

 

Caution锛?/span>

[1]. When the plasmid contains unstable fragments, it can reduce the probability of false recombination when cultured at 30 C. If we convert control pUC19 to calculate the transformation efficiency, it needs 37 degrees centigrade and 225 RPM revival for 60 minutes.

[2]. When the plasmid contains unstable fragments, it can reduce the probability of erroneous recombination when cultured at 30 C. If control pUC19 is used to transform the efficiency, it will need to incubate at 37 C for the night.