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What will students do in this lab?
What will students do in this lab?
Each student extracts DNA from 2 herbal supplements. This DNA will ultimately be sequenced to determine the contents of the supplements. They use 2 supplements to ensure that their centrifuge will be balanced, and to increase the chance that at least one of them will be successful. Students can use the supplements that we send them or use their own. Since supplements are expensive, most students use ours unless they already have a supplement at home that they use.
Note: Teas are OK, but they will need to be pulverized (in a blender or with a mortar and pestle).
Learning goals
Learning goals
You will extract DNA that is pure enough for PCR
You should safely and accurately use pipettors, centrifuge and a water bath
Safety Information
Safety Information
Everything in this lab is safe, but chemicals are lab grade, not food grade. Do not eat or drink. Do not catch things on fire. Remove any food, drink and dishes from the area before working.
Work on the provided bench paper to absorb any spills.
Centrifuge: Always balance your centrifuge, and wait until the centrifuge is done spinning before opening the lid.
Thermometer: The thermometers do not contain mercury. They are non-toxic, but they are breakable. If you break one, be careful of the sharp pieces, and dispose of it in your sharps container.
Everything used in this lab may be disposed of in the trash or drain.
Wash hands and clean counter tops when you are done.
MSDS: If you click through this link, you will find Material Safety Data Sheets (MSDS) for chemicals used in this lab. These are technical documents that contain safety information on all reagents for this lab. None of this information will be on homework or exams. However, you should read through the sections on "Hazards Identification," and "First Aid Measures"
Video: Preparing samples
Video: Preparing samples
Watch this video to learn how to prepare samples for DNA extraction.
If your sample is a capsule that you can pull apart, it probably contains powder and you are good to go.
If you are using a tea or something not ground up, you can grind it with a mortar and pestle, in a blender, etc. Make sure the mortar and pestle/blender/etc is very clean, or you might contaminate your sample with whatever you previously processed.
It is very important to use only a tiny amount of tissue. Aim for less than you see at 1:22 in the video, maybe half of that. Unfortunately, where I show the perfect amount of tissue (0.005g) at 1:35 , the tube is out of focus.
Video: How to use your centrifuge
Video: How to use your centrifuge
Watch this video to learn how to use your centrifuge.
The crucial things to remember when using a centrifuge:
Balance your centrifuge - you should have 2 tubes of equal weight opposite each other
Use the adaptors for the size tube you have. The biggest tubes, which you'll use in the DNA extraction, don't require adaptors. The other tubes do. You don't actually need to change the rotor, since there adaptors that work even for the tiny PCR tubes.
Video: DNA extraction step by step
Video: DNA extraction step by step
Watch this video to learn how to extract DNA from your supplements.
In this video, we carry out the whole protocol and give tips and tricks.
DNA extraction means that you remove all of the proteins and other cell contents to obtain purified DNA from all of the species present in your supplement.
Read this protocol carefully before beginning your DNA extraction.
Follow the protocol to extract DNA from 2 herbal supplements, either the ones we provided or supplements of your choice. The supplements need to contain plants, as we are sequencing plant DNA for species identification
Please take pictures as you carry out the lab so you can show that you did the lab.
Take this quiz before beginning your DNA extraction.
This is to ensure that you read the protocol before you begin.
You are encouraged to look at the protocol as you take this quiz
Record the names of the supplements from which you extracted DNA here
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