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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:
NMY51 chemically Yeast Express Competent Cells
- 库存:
货源充足
- 供应商:
LSM Bio
- 规格:
0.1ml*10
NMY51 Chemically Competent Cell
Packing 10 x 0.1 ml/tube
Store at –80°C
Genotype
MATa his3Δ200 trp1-901 leu2-3,112 ade2 LYS2::(lexAop)4-HIS3 ura3::(lexAop)8-lacZ ade2::(lexAop) 8-ADE2 GAL4
Alternative name
NMY51 chemically Competent Cells,NMY51 Competent Yeast,NMY51 Competent Cells, NMY51 chemically Yeast Express Competent Cells
Transformation Protocol
1.Please store NMY51 Chemically Competent Cell in -80°C for late use.
2. Thaw a tube of NMY51 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 30 minutes. Do not mix.
5. Heat shock at exactly 42°C for exactly 90 seconds and then immediately transfer the tube back into ice for 5min. Do not mix.
6. Place on ice for 5 minutes. Do not mix.
7. Add 800ul LB then put on the Constant temperature shaker (150 rpm) for 45min,Warm selection plates to 37°C.
8. Pellet the mixture by centrifugation at 4000rpm/min for 3min. Pour off about 200ul 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
Membrane‐Based Yeast Two‐Hybrid System to Detect Protein Interactions
1×YPAD liquid medium (see recipe ) NMY51 yeast strain: MATa his3Δ200 trp1‐901 leu2‐3,112 ade2 LYS2::(lexAop) 4 ‐HIS3 ura3::(lexAop) 8 ‐lacZ ade2::(lexAop) 8 ‐ADE
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
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