LAB REPORT ASAP 2PAGES

Write up the three operations: extraction, PCR and agarose gel.

 Your report should include the following:

· Name your test food ( Cheetos chips)

· List of contents of the wells

1- non GMO control, with DNA plant master mix.

2- Non GMO control, with DNA GMO master mix

3- Test food DNA, with plant master mix

4- Test food DNA, with GMO master mix

5- GMO positive control DNA, with plant master mix

6- GMO positive control DNA, with GMO master mix

7- PCR molecular

8- Empty

GMO Extraction

Introduction

Genetic engineering has been used to improve traits of many crops such as yield,

diseases resistance, pesticide tolerance. These crops are generally known as

genetically modified organisms or GMOs.

There are some who want to not consumer GMO crops for various reasons, so

being able to detect plants and foods that do not contain GMO is important

There are two ways to detect GMO

oUse an ELISA assay, which is an antibody-based test, to detect the protein

that are being expressed.

O Use PCR that can amplify engineered sequences such as

35S promotor from the cauliflower mosaic virus (CaMV 35S)

NOS (nopaline synthase) terminator from Agrobacterium tumefaciens

The PCR assay we will use primers for both NOS and CaMV 35S (red dye added) as well as primers against plant DNA (green dye) as a check on the extraction and

amplification.

Objective

Test crops or processed foods for genetic modification.

In this lab DNA will be extracted from food materials

Materials

· P-20 (2 to 20 ul)

· P-200 (20 to 200 ul)

· P-1000 (200-1000 ul)

· Distilled water

· Screw cap tubes

· InstaGene matrix

· Transfer pipettes

· Mortar and pestle – 1 for each group

· Heating block at 95C to 100C

· Microfuge tubes

· Microfuge

· Balance

· Weigh boats

· At least 1 test food

· corn meal, corn bread mix, taco shell, corn snacks etc

Procedure

Wear nitrile gloves

Label one screw cap tube with 500ul of Instagene matrix ‘non-gmo’ and the other ‘test’

Wash your mortar and pestle with detergent and dry.

First weigh out 1 gram of the non GMO food in a weigh boat.

Place the food in mortar and add 5 ml of the distilled water and grind for at

least 2 minutes until a slurry is formed.

Add another 5 ml of water and grind further until the slurry is smooth enough

to pipette.

Add 50ul of the ground slurry to the screw cap tube containing the InstaGene

matrix that is labelled ‘non-gmo’ You can use a transfer pipette – 50ul is the

lowest graduation.

Recap tube and shake well.

Wash the mortar and pestle with detergent and dry.

Repeat the above steps with the test material and place 50ul of the slurry in the test screw cap tube labelled test.

Place the non-gmo and test food screw cap tubes in a 95C heating block for 5

minutes.

Spin tubes in microfuge for 5 mins on maximum speed.

Store tubes on ice.

GMO PCR

Objective

· Test crops or processed foods for genetic modification

· In this lab DNA will be amplified

Materials

•Ice bath

GMO master mix – red

Plant master mix – green

•

GMO positive control

•

Test food DNA

•

PCR tubes

•

PCR adaptors

•

Foam micro tube holders

•

2-20ul micropipette

•

200 ul micropipette

•

Thermal cycler

Procedure

PCR set up

Prepare six PRC tubes (20 ul)

1- 20ul non GMO control, with DNA plant master mix.

2- Non GMO control, with DNA GMO master mix

3- Test food DNA, with plant master mix

4- Test food DNA, with GMO master mix

5- GMO positive control DNA, with plant master mix

6- GMO positive control DNA, with GMO master mix

7- PCR molecular

8- Empty

Thermal Cycler set up

· After the PCR has been completed the tubes will be frozen at -20C.

· Conclusions

· Name your test food:

( Cheetos chips)

· List of contents of the wells

9- non GMO control, with DNA plant master mix.

10- Non GMO control, with DNA GMO master mix

11- Test food DNA, with plant master mix

12- Test food DNA, with GMO master mix

13- GMO positive control DNA, with plant master mix

14- GMO positive control DNA, with GMO master mix

15- PCR molecular

16- Empty

· Was your unknown a GMO food? What is your evidence that makes you draw that conclusion.

· In regards to the controls what does a band in these lanes tell you

· 1

· 3

· 5

· Why do you not expect a band in lane 2

Results

The GMO food did not bind with plant primer. The result was negative