Tech Challenge 
Analysis of Gene Transformation Etoh Etc., a biofuels company, wants to determine the number of copies of a gene related to ABE fermentation that their transformed bacterial strains have acquired. There are a number of ways to quantify gene copies, but they want to be able to screen hundreds of transformed colonies easily, accurately, and quickly. Which of the following methods would be ideal for this company? - Southern blots performed with restriction enzyme-digested genomic DNA, using an Electrophoresis Blotting System.
- Standard PCR and electrophoresis of genomic DNA, using a High-Throughput Horizontal Gel System.
- Real-time PCR of genomic DNA, using the Techne PrimeQ Real-Time PCR System.
- Protein over-expression assessments by ELISA, using the Bio-Tek ELX 800 Universal Microplate Reader.
Answers:
- Southern blots performed with restriction enzyme-digested genomic DNA, using an Electrophoresis Blotting System.
Incorrect. This system is low-throughput, so it would not be able to process hundreds of samples per day unless they had dozens of these systems going at once.
- Standard PCR and electrophoresis of genomic DNA, using a High-Throughput Horizontal Gel System.
Incorrect. Although this system can process many samples at a time, standard PCR cannot accurately quantify the number of copies of any given gene. It is a qualitative analysis.
- Real-time PCR of genomic DNA, using the Techne PrimeQ Real-Time PCR System.
Correct! This system can analyze hundreds of colonies per day because it performs PCR on 96-well plates. The analysis software, Quansoft, comes with the unit, making analysis easy. And, measuring the level of PCR product at each step in the cycle creates an accurate PCR product profile from which original gene copy number can be inferred reliably.
- Protein over-expression assessments by ELISA, using the Bio-Tek ELX 800 Universal Microplate Reader.
Incorrect. This instrument is not designed for reading ELISA. The customer would have to purchase additional software to analyze ELISA plates, so it is not the easiest option.
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