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- LSM Bio
- CHC00032
- 2025年07月09日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
负80摄氏度
- 保质期：
6month
- 英文名：
M15 chemically E.coli Express Competent Cells
- 库存：
货源充足
- 供应商：
LSM Bio
- 规格：
0.1ml*10
M15 Chemically Competent Cell
Packing 10 x 0.1 ml/tube
Store at –80°C
Genotype
lac,ara,gal,mtl,recA+,uvr+[pREP4,lacI,kanaR]
Alternative name
M15 chemically Competent Cells ,M15[PREP4] chemically Competent Cells,M15[PREP4] Competent E.coli,M15[PREP4] Competent Cells, M15[PREP4] chemically E.coli Express Competent Cells
Discription
E. coli M15 DZ291
M15 Chemically Competent Cell includes the pREP4 plasmid which confers kanamycin resistance and constitutively expresses the lac repressor protein encoded by the lac I gene.
M15 cannot be infected by lambda phages
PL promoter introduced; however, is not active
Transformation Protocol
1.Please store M15 Chemically Competent Cell in -80°C for late use.
2. Thaw a tube of M15 Chemically Competent Cell锛?00μl 锛塷n ice .
3. Add 1µl plasmid DNA or 10µl ligation product to the cell mixture. Carefully flick the tube 4–5 times to mix cells and DNA. Do not vortex.
4.Place the mixture on ice for 25 minutes. Do not mix.
5. Heat shock at exactly 42°C for exactly 45 seconds and then immediately transfer the tube back into ice for 2min. Do not mix.
6. Place on ice for 2 minutes. Do not mix.
7. Add 700ul LB then put on the Constant temperature shaker (200 rpm) for 60min,Warm selection plates to 37°C.
8. 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.
9.Spread the cell on the agar plates by antibiotic, then place it on 37°C for 1hour.
10. Incubate overnight at 37°C.
Caution:
1.STORAGE AND HANDLING: Competent cells should be stored at
–80°C. Storage at –20°C will result in a significant decrease in transformation efficiency. Cells lose efficiency whenever they are warmed above –80°C, even if they do not thaw.
2. This product is FOR RESEARCH USE ONLY!
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文献和实验a chilled, sterile pipette tip to transfer the competent cells to chilled, sterile 17 x 100-mm polypropylene tubes. Store the cells on ice. Glass tubes should not be used since they lower the efficiency of transformation by ~10-fold
,etc.).If a cooling shaker is not available- grow the cells at room temperature.More discussions on making competent cell as well as references can be found in TIBS articles "Preparing μltra-competent E.coli" and "Better competent cells"It is also possible
;ltra-competent E.coli" and "Better competent cells" It is also possible to transform cells straight from plate.It is convenient but you should expect low efficiency.See the following reference for more details (as well as more information
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