Main Entry: re·com·bi·na·tion Pronunciation: \ˌrē-ˌkäm-bə-ˈnā-shən\ Function: noun : the formation by the processes of crossing-over and independent assortment of new combinations of genes in progeny that did not occur in the parents
InstaGene™ matrix: What Function Does It Perform? InstaGene matrix consists of a suspension of negatively charged, microscopic beads that bind divalent cations such as magnesium (Mg2+). It is important to remove divalent cations from the extracted DNA samples because the cations assist enzymes that degrade the DNA template. When cheek cells are lysed and boiled in the presence of InstaGene matrix, the divalent cations released from the cells bind to the beads, and the heat inactivates the DNA-degrading enzymes. The beads are pelleted by centrifugation, and the supernatant, which contains clean, intact genomic DNA, can be used as template in PCR reactions. The beads in the InstaGene matrix quickly settle out of the suspension. It is therefore extremely important that the vial of matrix be thoroughly mixed before pipetting aliquots for each student workstation, so that the aliquots contain equivalent amounts of beads. If the DNA samples are not going to be amplified within 24 hours, they can be stored in the refrigerator in the InstaGene matrix for up to 1 week. For longer storage, place samples in the freezer to prevent DNA degradation. Before the samples are used in PCR, the beads should be repelleted by centrifugation just prior to making up the PCR reactions.
Master Mix: What Is It? The master mix contains a mixture of nucleotides, or dNTPs (dATP, dTTP, dCTP, and dGTP), buffer, and Taq DNA polymerase. Complete master mix is prepared by adding primers to the master mix just prior to the laboratory period. When 20 μl of the DNA template is added to 20 μl of complete master mix, all of the necessary components for a 40 μl PCR reaction are present. Note: Once the master mix and primers are mixed, the complete mix should be kept on ice and used within 30 minutes to 1 hr. These reagents are extremely sensitive. Why Are the Primers Red and Green? The primer mixes contain PCR-compatible dyes that allow students to easily visualize and distinguish the different master mixes. The dyes also migrate in the gel giving a visual demonstration of electrophoresis.
Investigation of genetic modification in maize and soymilk
Genes to identify: 35S promoter, NOS terminator, and
a) DNA Extraction
b) DNA Amplification
c) Electrophoresis of DNA
Analysis of Results
GMO’s are organisms that have been modified through
In the United States, under guidelines issued by USDA's
Animal and Plant Health Inspection Service, genetic
engineering is defined as the genetic modification of
organisms by recombinant DNA techniques (USDA, 2007)
According to the USDA, crops have been genetically
modified ever since 1996.
These crops have all kind of modifications, such as:
herbicide tolerance, pesticide tolerance, and bacterial
Steps for genetic
modification of a
a) First, find the
ideal protein to
b) Isolation of the
gene of interest.
c) Insertion of the
d) Back cross to
35S promoter from the cauliflower mosaic
virus (CaMV 35S)
Nopaline synthase (NOS) terminator from
Photosystem II (PSII)
Our hypothesis is that the genes 35S and NOS
will appear in the gel. The 35S gene will be
found at 203bp and NOS at 225bp. We
believe the corn will be identified as GMO
while the soymilk won’t. In addition, both the
corn chips and the papaya will be indentified
a) Weighing and
Grinding of samples
b) Pipette 50µL of Slurry
into tube containing
c) Flick tube and place
in a 95 degrees
celsius water bath for
d) Place tubes in
microcentrifuge for 5
Non-GMO Test food Test food
a) 20µL of indicated
master mix is
dispensed to each
of the samples.
b) 20µL of DNA
dispensed to their
c) Once the samples
tubes are put in
the thermal cycler.
GMO (+) PMM
GMO (+) GMM
PMM= Plant molecular marker
GMM= Genetic molecular
a) The gel used was a 3% Agarose
b) To prepare for electrophoresis,
10µL of Orange Loading dye was
deposited into each of the
c) 20µL of Molecular weight
ruler(mwr) was deposited to wells
to wells 1 and 10.
d) 20µL of samples were dispensed
to wells 2-9.
e) Electrophoresis was run for
30min. at 100V.
f) After electrophoresis, the gel was
stained with ethidium bromide
pads for 20min.
1 Molecular weight Ruler
2 Non-GMO PMM
3 Non-GMO GMM
4 GMO (+) PMM
5 GMO (+) GMM
6 Test PMM
7 Test GMM
8 Test PMM
9 Test GMM
10 Molecular Weight Ruler
One can conclude that the maize was
genetically modified, while the soymilk,
papaya, and corn chips are inconclusive.
The experiment must be repeated because
the results were affected by human errors,
such as bad pipetting technique.