Chloroplast subclassification experiments

Summary

This experiment is based on the "Guide to Cellular Experiments", translated by Huang Peitang et al.

Operation method

Chloroplast subclassification

Materials and Instruments

Chloroplast
Lysis buffer
Microcentrifuge Tubes Mini Centrifuge

Move

1. Pipette a chloroplast suspension containing 1 mg of chlorophyll into a microcentrifuge tube.

2. Centrifuge in a small centrifuge at 14000 r/min for 30 seconds and discard the supernatant.

3. Add 1 ml of Lysis Buffer, shake, and ice bath for 5 minutes.

Lysis Buffer:

10 mmol/L HEPES-KOH, pH 8.0

4. Centrifuge at 14000 r/min for 2 min in a minicentrifuge and discard the supernatant (matrix-rich fraction).

5. Wash the membrane components with 1x SH at least twice (enriched with vesicle fractions).

Preservation Solution:

20 mmol/L HEPES-KOH, pH 8.0

20 mmol/L sorbitol

125 mmol/L KCl

Contains 30% (v/v) DMSO or 30% ethylene glycol

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