main objective of GMO detection experiment is extract DNA from food material and to test genetic modification on processed food.

GMO Detection

Lab report #3

Objective

The main objective of GMO detection experiment is extract DNA from food material and to test genetic modification on processed food.

Background

Genetically modified food is very common nowadays and over 60% of food products in United States are genetically modified. To modify a product genetically means to alter the DNA in a way which is not natural. This is usually through genetic engineering which allow for the introduction of characteristics as well as having control over characteristics in comparison to original traits such as mutation breeding and selective breeding. The process of producing genetically modified food started back in 1994 by Calgene through a genetically modified tomato (Flavr Savr) though it was unsuccessful that year (Huang et, al, 2013). Since then most genetic engineers have focused mainly on cash crops that are of high demands to farmers that can resist herbicides, pathogens and have better nutrients constituents. Some of such food products include; soybeans, papaya, Canola, corn and cotton. However, due to increased production of genetically modified foods, the demand for GM testing in food products has increased to meet the demand of the consumers for both who want and those do not want genetically modified food products. The two-main method applied by scientist has been protein-based lateral flow strip tests or ELISA test and DNA-based polymerase chain reaction (PCR). Strip test detects a specific protein produced by GM products genetically modified DNA while PCR tests analysis the DNA directly. The current lab test however was used to test for genetic modification of crops or processed food.

Procedure

With the nitrile gloves on, one screw cap tube was labeled with 500ul Instagene matrix ‘non-GMO’ and the other on ‘test’. Then, the mortar and pestle were washed with detergent and dried. A weigh boat was used to first measure 1 gram of non-GMO and the food was placed in the mortar. Distilled water measuring 5ml was added in the mortar and grinding was done for around 2 minutes until slurry formed. Five ml distilled water was added into the slurry and was further grinded until it was smooth enough to pipette. In the screw cap labeled ‘non- GMO’ containing InstaGene matrix, 50ul of ground slurry was added using a pipette. The tube was recapped well and was mixed well through shaking the tube. The mortar was washed well with detergent and dried. The above procedure was repeated with the test material but 50ul slurry was added in a screw cap labeled ‘test’. The tubes were further spanned in a microfuge for 5 minutes on a maximum speed and after were stored on ice.

GMO PCR Procedure

PCR set up

Six PCR tubes (20ul) were prepared.

List of the appropriate tube numbers, primers and DNA samples

The first lane tube had 20ul non-GMO control, with DNA plant master mix. Second lane non-GMO control, with DNA GMO master mix, third Test food DNA, with plant master mix, fourth Test food DNA, with GMO master mix, fifth GMO positive control DNA, with plant master mix, sixth GMO positive control DNA, with GMO master mix, seventh PCR molecular and eighth empty. Finally, the tubes were frozen at -20 ° C.

Table 1. Thermal Cycler set up

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Results

The GMO food did bind with plant primer. In addition, it was a GMO food because in lane 5 a 200 bp band was present which means the food contained the GMOs.

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Figure 1. DNA ladder in gel electrophoresis.

Conclusion

1. Name your test food.

Cheetos chips.

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

Yes, it was a GMO food because in lane 5 a 200 bp band was present which means the food contained the GMOs.

3. In regard to the controls what does a band in these lanes tell you.

Lane 1, Lane 3 in these two lanes there is a band which means the food is GMO positive. In Lane 5 the absence or presence of the 200pb band indicates if the food tests have GMOs or not.

4. Why do you not expect a band in lane 2?

Lane 2 has non-GMO food control with GMO primer. That’s why it will not band.

RFERENCES

Huang, H., Cheng, F., Wang, R., Zhang, D., Yang, L. (2013) . “Evaluation of Four Endogenous Reference Genes and Their Real-Time PCR Assays for Common Wheat Quantification in GMOs Detection.” PLOS One 8.9

 

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