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Wet Lab Overview

Our target gene is the SLCO1B1 located on chromosome 12. In humans, encodes a protein transporter OATP (organic anion transporter protein) that works to move statins into tissues. OATP1B1 moves simvastatin and the other statins into the liver, the site of their action, to inhibit the HMG GoA reductase, resulting in decreased production- level of cholesterol. Also, the liver is the main place where statins are metabolized. If reduced levels of simvastatin are transported into the liver, there is a high risk of severe adverse effects such as myopathy. People who are homozygous for the C allele of the rs4149056 may appear drug-toxicity and ADRs, such as muscle’s pain including myopathy, when simvastatin is administered.

Ιn order to find out each patient’s genotype, we performed a test with the BentoLab, a Portable Molecular Biology Laboratory. It is worth noticing that the particular portable laboratory we are using has already been introduced by a previous iGEM team, iGEM UCL_E 2013. The UCL team introduced the portable laboratory called BentoLab back at the iGEM 2013 competition. Even though we use this BentoLab for obvious and traditional reasons, we tried to combine its cause with beneficial software technology. After taking the test’s results, Artificial Intelligence software, which we created, undertakes to instruct the physician on the dose to be taken by his patient.

The following steps to get a complete test with the BentoLab and get the final result that will be analyzed later by Artificial Intelligence are presented below:

  1. BentoLab Specifications
  2. Selecting the blood samples.
  3. DNA extraction from the whole blood samples using DNA extraction Kit.
  4. Nanodrop
  5. PCR Allele - specific
  6. Primer designing for Allele- Specific PCR
  7. Standardization of the PCR allele-specific based on samples with known genotype
  8. PCR-Allele specific for the analysed samples (samples with unknown genotype)
  9. Amplification
  10. Electrophoresis
  11. Results – Blue LED light

Working on Project Hippocrates, we made day-to-day progress that it is captured in our Team’s Notebook for the WetLab called LabBook.

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