Lab 5: Biochemistry III

Question 1

learning objectives

Answer 1

Gel Electrophoresis of Amplicons (PCR part 2)
• Describe and/or sketch the process of agarose gel electrophoresis (how the gel tank
setup allows the separation of DNA molecules by size) and the role of each reagent
(reagents include SYBR Safe, loading dye (bromophenol blue, xylene cyanol, &
glycerol), agarose, 1× TAE buffer, DNA molecular weight standard, and UV light).

• Be able to prepare all reagents necessary for agarose gel electrophoresis: % agarose,
1x buffer from a stock, and dilution of loading dye.

• Estimate the size of DNA fragments using a DNA molecular weight standard.

• Determine the outcome of a GMO detection procedure by PCR amplification using
plant-specific and GMO specific primers, interpret AGE results from such
amplifications, and troubleshoot errors.

Question 2

How would you genetically engineer a plant

Answer 2

Plasmid Vector:
• Antibiotic gene (select bacteria with
plasmid vector)

• nos gene (to increase infectivity)

• CaMV Promotor (to increase RNA
transcription in plants)

• Pesticide gene (endotoxin, when
expressed by plant, kills pest)

Bacteria that infects plant:
• Agrobacterium tumefaciens

Photosystem II gene found in plants:
• PSII (psbA chloroplast gene)