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BIT 410/510: Manipulating Recombinant DNA

Overview:

  • Experience biotechnology research and explore its impact in research and industry.
  • Students will gain hands-on experience with modern biotechnolgy techniques and develop technical skills for future laboratory research and industry postions.
CHO GLO cells after receiving 10 uL 100 mM ATP. Lionheart FX was used to capture images. The video is at 2X speed. ATP diffuses from left to right and the fluorescence drops after the ATP reaches the cell. Video and experiment performed by BIT alumni, Rose Krebs.

Lectures:

  • What is DNA Cloning?
  • Experimental Design
  • PCR Cloning and Gibson Assembly
  • Screening Recombinant Clones
  • Animal Cell Culture and Aseptic Technique
  • Next Generation Sequencing
  • RNA Interference and Genome Editing with CRISPR-Cas9

Labs:

CaMPARI Cloning

  • Hypothesis Design
  • Cloning by PCR
  • Plasmid DNA Purification/Quantification
  • Ligation and Transformation
  • Screening Clones (PCR, Sequencing)
  • Gibson Assembly

CaMPARI Expression in E. coli and animal cells

  • Hypothesis Testing
  • Protein Purification
  • SDS-PAGE and Western Blotting
  • Animal Cell Culture and Transfection
  • Fluorescence Microscopy
  • Quantitative Data Anaylsis

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