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Lab 6 – Colony PCR Screening and Sanger Sequence Alignment

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This lab involves screening transformed E. coli colonies for the presence of the CaMPARI insert using colony PCR and gel electrophoresis. It also includes analysis of Sanger sequencing data to confirm insert identity and orientation using SnapGene. Students identify errors such as truncations, SNPs, or misalignments that may affect downstream expression.

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Sequence Alignment with SnapGen
❖ Data for activity
❖ pHLsec-CaMPARI reference located on Moodle under “Important
Links”
❖ Sanger Sequencing data located in Lab 6 Google Folder

❖ Do you find any mismatches? Where are they located on the
reference sequence?
❖ Would this cause issues when the gene is expressed?

Primer Region of Quality Sequence on plasmid Misaligned nucleotides

pCAG-F [insert text here] [insert text here]

Bglob-pA R [insert text here] [insert text here]

, BIT 410/510 Lab 6
PCR Screening Gel &
Sanger Sequence Alignment with SnapGene

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Institution: North Carolina State University
Course: BIT 410

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Uploaded on: March 30, 2025
Number of pages: 15
Written in: 2023/2024
Type: Class notes
Professor(s): Bullock
Contains: All classes

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Lab 6 – Colony PCR Screening and Sanger Sequence Alignment

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