GMO Lab report (1 page) 6 hours ASAP

Biol 390-Lab 12 GMO Extraction

Objective

· Test crops or processed foods for genetic modification.

· In this lab DNA will be extracted from food materials

Background

· 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

· Use an ELISA assay, which is an antibody-based test, to detect the protein that are being expressed.

· 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.

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 95oC to 100oC

· Microfuge tubes

· Microfuge

· Balance

· Weigh boats

· At least 1 test food

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

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Procedure

Prep before lab

· Add 500ul of InstaGene matrix to each of 8 screw cap tubes

· Each group gets 2 tubes

DNA extraction

· 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 95oC heating block for 5 minutes.

· Spin tubes in microfuge for 5 mins on maximum speed.

· Store tubes on ice.