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Lab #5 – Gibson Assembly, Transformation, and Interim Lab

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In this lab, we used three protocols: Gibson Assembly, Transformation, and the Interim Lab. Each of these protocols was critical for progressing our CaMPARI project: Gibson Assembly was used to combine our linear DNA fragments into a complete plasmid (pHLsec-CaMPARI). Transformation allowed us to introduce this newly assembled plasmid into competent E. coli DH5-alpha cells. The Interim Lab helped us track our progress and ensure our plasmid was constructed correctly before proceeding to mammalian expression. These protocols are essential steps in constructing a functional plasmid that can later be used to express CaMPARI in mammalian cells.

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Institution
North Carolina State University
Course
BIT 410










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Institution
North Carolina State University
Course
BIT 410
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Uploaded on
March 30, 2025
Number of pages
17
Written in
2023/2024
Type
Class notes
Professor(s)
Bullock
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All classes

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BIT 410/510
Lab #5
Gibson Assembly
Transformation
& Plate Art
Interim Lab

, Lab 5 Order of Operations
❖ In the Lab Activities Today:
❖ Activity 1 - Gibson Assembly and Transformation of E. coli
❖ Activity 2 - Plate Art
❖ Tomorrow(next business day):
❖ Interim Lab Materials
❖ You will receive photos of your plates to count colonies from.
❖ Please make sure to label your plates as instructed!
❖ This protocol should be included in your Lab 5 ELN submission!
❖ The video explains count transformants and inoculation → you
only have to worry about count transformants

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