Write up the three operations: extraction, PCR and agarose gel.
Your report should include the following:
Name your test food: Cheetos chip
List of contents of the wells
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
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