GMO LABORATORY PROCEDURE
DAY 1: ISOLATE DNA FROM SOY AND FOOD PRODUCTS
TODAY’s STARTING POINT
TODAY’s ENDING POINT
Please note: Each group of two will select EITHER wildtype or Roundup Ready soy OR a food
product. Note the product you are working with_______________________________________
and be sure to label all tube with your assigned lab number_______________________________
The protocol below is written for each group to analyze two samples (soy and food).
1.
TUBE #1: SOY LEAVES
Cut two pieces of soy leaf 1/4 inch in diameter. Put both pieces into
a 1.5 ml tube. Label with soybean type and your group number.
TUBE #2: FOOD
Crush a small amount of food on a piece of paper. Put crushed
food into a 1.5 ml tube about halfway to the 0.1 ml mark. Label with
the food type and your group number.
2.
Add 100 l of Edward’s buffer to each tube.
3.
Forcefully grind the plant tissue or food product for 1 minute.
4.
Add 900 l of Edward's buffer to each tube.
5.
Flick the tubes for 5 seconds to mix things up a bit.
6.
Place boiling caps on samples. Boil the samples for 5 minutes
in the heat block.
7.
Centrifuge the samples for 2 minutes to pellet cell and food
debris.
8.
Put 350 l of each supernatant into new tubes.
Labe the new tubes carefully and throw out the old tubes so you
don’t get confused.
9.
Add 400 l of isopropanol to each tube. Mix by turning the tubes
upside-down several times. Once mixed, leave the tubes at room
temperature for 3 minutes.
10.
Centrifuge for 5 minutes. Place tubes in the centrifuge with the
cap hinges pointing up.
11.
Pour off the supernatant from each tube.
12.
Completely remove the remaining liquid with a pipette set at 100
l.
13.
Air dry the pellets for 10 minutes.
14.
Add 100 l of distilled water to each tube.
15.
Pipette the liquid in and out. Try to wash down the side of the tube
underneath the hinge; this is where the DNA is. Let the tubes sit
for 5 minutes.
16.
Centrifuge the tubes for 1 minute.
17.
Store your samples in the freezer. Make sure the tubes are still
clearly labeled.
DAY 2: AMPLIFY DNA BY PCR
35S PROMOTER: Only found in genetically modified foods
Please note: Each group will set up one PCR reaction for 35S. Please label tube with your group
number and 35S.
35S PCR TUBE #1: SOY DNA
Label PCR tube with your group number and 35S SOY DNA (RR or NRR)
Example: M2 5 35S NRR
35S PCR TUBE #2: FOOD DNA
Label PCR tube with your group number and 35S FOOD DNA (F).
Example: M2 5 35S F
Add 23 l of the 35S primer/loading dye mix to each
tube. Allow 3 minutes for the PCR bead to dissolve.
Add 3.0 l of FOOD DNA (from Part I) to the PCR tube
labeled “FOOD DNA”
Add 3.0 l of SOY DNA (from Part I) to the PCR tube
labeled “SOY DNA”
TUBULIN: Found in everything
Please note: Each group will set up one PCR reaction for Tubulin. Please label tube with your group
number and T for Tubulin.
Tubulin PCR TUBE #1: SOY DNA
Label PCR tube with your group number and Tubulin SOY DNA (RR or NRR)
Example: M2 5 Tub NRR
Tubulin PCR TUBE #2: FOOD DNA
Label PCR tube with your group number and Tubulin FOOD DNA (F).
Example: M2 5 Tub F
Add 23.0 l of the Tubulin primer/loading dye mix to
each tube. Allow 3 minutes for the PCR bead to
dissolve.
Add 3.0 l of FOOD DNA (from Part I) to the PCR tube
labeled “FOOD DNA”
Add 3.0 l of SOY DNA (from Part I) to the PCR tube
labeled “SOY DNA”
Bring all four PCR tubes to the thermocycler or store
on ice (up to 24 hours) until ready.