Mitochondrial Morphology in Budding Yeast- W303

Published: 23-04-2017| Version 1 | DOI: 10.17632/2td7b68d9f.1
Contributor:
Shao-Ting Chiu

Description

Strain: W303a (873) Growth medium: YPD Fluorescence: MitoTracker Red Microscope: LSM780

Files

Steps to reproduce

實驗流程 Sample Preparation: • 873 vs. 873q2 in YPD: Mitotracker 能不能染 petite • 873 in YPD vs. 873 in YPG: Confocal 能不能分辨 morphology 的不同 • 一倍濃度與十倍濃度: 假設 stock solution 1mM (working concentration: 25-500nM) 1. Culture 873(W303a) and 873-q2(petite from W303a) in YPD; Culture 873 in YPG Genotype Growth Medium WT(W303) YPD WT(W303) YPG Rho0 YPD 2. Growth overnight; 3. Refresh for three hours 4. 量 OD. Collect ??體積 5. Wash for twice 1) Centrifuge 3000g 30sec 2) Remove the medium 3) Resuspend with H20 6. Centrifuge 3000g 30sec 7. Remove H20 8. Add 600 staining solution/ tube1 共六管 9. Incubate for 15 minute (30°C) 10. Wash with PBS 11. 取 1.5 ul 到玻片上 (*不可以壓玻片; 先用實驗室顯微鏡看過) 1) 兩個 sample /玻片; 不同濃度放一起