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- CHC00060
- 2025年07月16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：
KM71 chemically Yeast Express Competent Cells
- 库存：
货源充足
- 供应商：
LSM Bio
- 规格：
0.1ml*10
KM71 Chemically Competent Cell
Packing 100 x 0.1 ml/tube
Store at –80°C
Genotype
arg4 his4 aox1::ARG4
Alternative name
KM71 chemically Competent Cells,KM71 Competent Yeast,KM71 Competent Cells, KM71 chemically Yeast Express Competent Cells
Transformation Protocol
1.Please store KM71 Chemically Competent Cell in -80°C for late use.
2. Thaw a tube of KM71 Chemically Competent Cell锛?00μl 锛塷n ice.
3. Heat shock Carrier DNA at 95-100°C then immediately transfer the tube back into ice for 5min,2times.
4. Add 2-5µl plasmid DNA, Carrier DNA 10ul and PEG/LiAc 500ul to the tube. Carefully flick the tube 4–5 times to mix cells and DNA.
5.Place the mixture in the water 30°C for 15 minutes. (Flip it 6-8times at 7.5min)
6. Place the EP Tube in the water 42°C for 15 minutes. (Flip it 6-8times at 7.5min)
7. Pellet the mixture by centrifugation at 500rpm/min for 40s. Pour off cell supernatant, then resuspend the cell pellet in the ddH2O 400ul..
8. Pellet the mixture by centrifugation for 30sec., Pour off cell supernatant. resuspend the cell pellet in the ddH2O 400ul.
9.Spread the cell on the agar plates by antibiotic, then place it on 29°C for 48-96hr.
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
J-6B centrifuge)for 10 min.at 4℃.4度2500g离心10分钟。5、Resuspend cells gently in 80 ml of ice cold "TB".小心用80ml预冷TB重悬细胞。6、On ice for 10 minutes.(30min)菌液置冰上10分钟。7、Spin at 2500 x g (5000 rpm in a Sorvall GSA,5500 rpm in a Sorvall SS-34,or 3000 rpm
! pH 6.7 - 7.0 Note: Competent Cell s are fragile (cell wall is thought to be weakened),therefore treat the cells gently when preparing these cells.Do not vortex or pippette up and down to resuspend the cells.Do not spin
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