Week 10 HW: Week 10 — Advanced Imaging & Measurement Technology

Homework: Final Project

(Measurement Draft)

The main aspect of my project that I will measure is the functional activity of a mutated InaZ construct, specifically whether it increases ice nucleation efficiency relative to a control InaZ construct. I will perform this measurement using a controlled freezing assay in which replicate samples are cooled gradually and monitored for the onset of ice formation. The primary data collected will be the temperature at which freezing begins in each sample. Ice formation will be detected through optical observation of crystal formation and through temperature sensors that record the freezing point. In addition, I would verify the identity of the mutated construct using PCR, gel electrophoresis, and DNA sequencing. PCR and gel electrophoresis would be used to confirm the presence and approximate size of the inserted DNA, while DNA sequencing would confirm that the engineered inaZ mutation is correct. Together, these measurements allow me to confirm both that the construct was built properly and that it produces the intended increase in ice nucleation activity.

Restated:

Functional Assay: Ice Nucleation

Objective: Measure efficiency relative to a control construct.

Method: Controlled freezing assay with gradual cooling of replicate samples.

Data Points: Freezing onset temperature, recorded via optical observation and thermal sensors.

Genetic Verification

PCR & Gel Electrophoresis: Confirm the presence and approximate size of the inserted DNA.

DNA Sequencing: Verify the exact sequence of the engineered inaZ mutation.

Homework: Waters Part I — Molecular Weight

Work to be Added

Homework: Waters Part II — Secondary/Tertiary structure

Work to be Added

Homework: Waters Part III — Peptide Mapping - primary structure

Work to be Added

Homework: Waters Part IV — Oligomers

Work to be Added

Homework: Waters Part V — Did I make GFP?

Work to be Added