엽록체 분리

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엽록체 분리

목차

Exp6. 엽록체 분리
1. 실험 목적
2. 실험 재료
3. 실험 방법

Exp7.아세톤에 의한 단백질의 침전
1. 실험 원리
2. 실험 재료
3. 실험 방법
4. 실험 결과
5. 고찰
6. 참고문헌

본문내용

2. 실험 재료
시금치, isolation buffer, blender, 냉각 원심분리기, e-tube, pipette

3. 실험 방법
1) Weigh out 40 g of fresh spinach (Spinacia oleracea) leaves from which the major veins have been removed.

2) Tear the leaves into small pieces and place them in a blender with 200 mls of cold isolation buffer (= 0.3 M sorbitol; 0.1 M Tris-Cl, pH 7.8; 5 mM MgCl2; 10 mM NaCl).

3) Blend the mixture at low speed for 10 seconds. Use a glass rod to push any tissue pieces down the sides of the jar into the solution, and blend for 10 more seconds.

4) Filter the homogenate (called a tissue `brei`) through 8 layers of cheesecloth into a beaker. Add approximately 40 ml of filtrate to each of four 40 ml centrifuge tubes, making sure they are balanced (+ 0.1 g).

5) Centrifuge at 4oC for 5 minutes at 1000 x g .

6) Decant and discard the supernatant from all 4 tubes. Add 0.5 mls of cold isolation buffer each tube and gently resuspend the chloroplasts with a paint brush.

7) Store the centrifuge tube on ice, away from bright light.

참고 자료

http://dragon.seowon.ac.kr/~bioedu/bio/ch04.htm
http://www.cheric.org/board/view.php?code=f03&seq=11354&page=17
http://cook.hyejeon.ac.kr/html/reportboard/uploadfile/%EB%8B%A8%20%EB%B0%B1%20%EC%A7%8 8.hwp