Haynes Lab:Notebook:Shared/Cloning Bootcamp

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Project Description/Abstract

This is the shared Notebook for Haynes Lab Cloning Bootcamp, a molecular biology training program organized by Dr. Karmella Haynes at Arizona State University.



STAGE A: Curating the Haynes Lab Plasmid DNA Collection

You have been assigned a plasmid or plasmids to “miniprep,” “verify,” and “submit to the Registry.” What does this mean and how do you get started? How do you finish? This guide will help you to complete your tasks, and learn about DNA and synthetic biology along the way. The purpose of curating the plasmid collection is to take some DNA from our freezer, produce a large batch of it, confirm that the reproduced DNA is not damaged, and to send that batch of DNA to a repository where the DNA will be shared with the world. This is a very important and educational task.

High-level concepts that you will learn about in greater detail include...

  • Synthetic biology - Using DNA parts or modules (like the ones in the Haynes Lab Plasmid Collection) in novel combinations to create artificial behaviors in living cells such as glowing, changing color, attacking tumors, eating up pollutants, computing, etc.
  • Cloning - The process of extracting a defined fragment of DNA from a larger source (such as a chromosome) and using it for basic research, or making “recombinant” (hybrid) pieces for synthetic biology.
  • Bacteria as workhorses - DNA is so small that it is hard to manipulate directly, so scientists use bacteria as living containers for storing and making copies of cloned DNA.

STAGE A consists of four Sessions:

  • SESSION 1 - Transforming Cells With Plasmid DNA
  • SESSION 2 - Growing Liquid Cultures of Transformed Cells
  • SESSION 3 - Plasmid Extraction and Validation
  • SESSION 4 - Reading DNA Sequencing Data