DNA Extraction from wheat germ Salk Institute for DNA提取小麦胚芽索尔克研究所.pptVIP

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2017-02-13 发布于上海
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DNA Extraction from wheat germ Salk Institute for DNA提取小麦胚芽索尔克研究所.ppt

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* Salk Institute Mobile Lab DNA Extraction from wheat germ Protocol Salk Institute Mobile Lab Step #1 Obtain a numbered 50 ml conical vial with 10 ml of water. Write your number here:________ Step #2 To the tube with 10 ml water add 1g (1 pinch) of raw wheat germ. The wheat germ should settle to fill 1/2 the cone Step #3 Cap the tube and vigorously shake to mix for about 30 seconds. What do you think the shaking is doing to the wheat cells? After shaking - let the cell walls settle to the bottom What’s in the cloudy solution? Where is the DNA at this point? Step #4 Using a transfer pipet add 3 ml of 25% dish soap to the tube with water and wheat germ. Steps #5-6 Cap the tube and slowly invert several times to mix (DO NOT SHAKE). Loosen the cap slightly before placing tube in the hot water bath. Heat at 65°C for 5 minutes. What do you think the soap will do? How will the hot water bath help? Bonus question: Why does washing your hands with soap and hot water kill bacteria on your skin? Step #7 While the tubes are heating label a 1.7 ml eppendorf tube with your initials. Carefully open the epi tube and place it into the rack Steps #8-9 Before removing your tube from the 65°C water fill a transfer pipet with 3ml of alcohol. Bring the 50 ml conical tube back to your table and as soon as possible add the 3ml of alcohol to the tube. Step #10 Watch the DNA rise out of the green soap layer and into the alcohol layer. What color is DNA?? What color do you think YOUR DNA is? Why? Step #11 After waiting a few moments to allow the DNA to precipitate remove the DNA using an inoculation loop and place in your eppendorf tube. Try to avoid any wheat germ debris that is present. Step #11 cont. Spinning the loop will help get the DNA off Make sure to close the epi tube securely (you should hear it “click”) Congratulations! You have just done something that scientists do everyday in the lab! Your DNA will last you for years – it’s very stable when stored in isopropanol