Shreffler:Notebook/retinoic acid trans signaling/2009/06/02

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Concentrating DNA:

(Note: We will use Sample 2x, which has a concentration of 112.3 ng/ml and a volume of 540 ul)

1. Split the volume into 2 tubes = 270 in each epp. tubes

2. Add 3 ul of glycogan to each tube

3. Add equal volumes of isopropanol to each tube (total of 30 ul of isopropanol to each tube)

4. Incubate for 30 min at room temperature

5. Spin for 15 min at max speed

6. Remove the liquid carefully without disturbing the pellet with a pipet and place into another tube (Keep liquid in case your steps need to be retraced)

7. Wash the pellet with 75% ethanol (100ul) and spin for 3 min at max speed

8. Remove the liquid carefully without disturbing the pellet with a pipet and place into another tube (Keep liquid in case your steps need to be retraced)

9. Repeat wash

10. Remove the liquid carefully without disturbing the pellet with a pipet and place into another tube (Keep liquid in case your steps need to be retraced)

11. Add 100% ethanol (100 ul) and spin for 2 min at max speed

12. Remove the liquid carefully without disturbing the pellet with a pipet and place into another tube (Keep liquid in case your steps need to be retraced)

13. Allow to dry at room temperature for 5 min, until the pellet turns transparent

14. Dilute each with 25 ul of sterile filtered water, then combine together = 50 ul concentrated DNA




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