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GMO Lab Report | Online Assignment Help

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

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 Your report should include the following:

Name your test food: Cheetos chip

List of contents of the wells

Conclusions

Was your unknown a GMO food? Corn

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

Biol 390-Lab 13 GMO – PCR

 

Objective · Test crops or processed foods for genetic modification

· In this lab DNA will be amplified

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 as a check on the extraction and amplification.

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

______

 

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

_______________________________________________

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.

Maryville University

Maryville University

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