The "Frankenplasmid" lab: An investigative exercise for teaching recombinant DNA methods

Derek M. Dean, Jason A Wilder

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We describe an investigative laboratory module designed to give college undergraduates strong practical and theoretical experience with recombinant DNA methods within 3 weeks. After deducing restriction enzyme maps for two different plasmids, students ligate the plasmids together in the same reaction, transform E. coli with this mixture of ligated DNA, and plate the cells on media that specifically select for hybrid plasmids. The main goal of the assignment is for students to deduce the gene map of one hybrid "Frankenplasmid" using the LacZ phenotype of its transformants, PCR, and restriction mapping. Our protocol results in a number of possible outcomes, meaning that students are mapping truly unknown plasmids. The open-ended nature of this assignment results in an effective module that teaches recombinant DNA procedures while engaging students with its investigative approach, increasing complexity, and puzzle-like quality. Moreover, the modular design of the activity allows it to be adapted to a more limited schedule, introductory courses, or more advanced courses.

Original languageEnglish (US)
Pages (from-to)376-383
Number of pages8
JournalBiochemistry and molecular biology education : a bimonthly publication of the International Union of Biochemistry and Molecular Biology
Volume39
Issue number5
DOIs
StatePublished - Sep 2011

Fingerprint

Recombinant DNA
Teaching
Plasmids
Exercise
Students
Restriction Mapping
Escherichia coli
Appointments and Schedules
Genes
Phenotype
Polymerase Chain Reaction
DNA
Enzymes

Keywords

  • PCR
  • Plasmids
  • Recombinant DNA
  • Restriction mapping

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

@article{a1511a912aff4dd4be5500bea474c00c,
title = "The {"}Frankenplasmid{"} lab: An investigative exercise for teaching recombinant DNA methods",
abstract = "We describe an investigative laboratory module designed to give college undergraduates strong practical and theoretical experience with recombinant DNA methods within 3 weeks. After deducing restriction enzyme maps for two different plasmids, students ligate the plasmids together in the same reaction, transform E. coli with this mixture of ligated DNA, and plate the cells on media that specifically select for hybrid plasmids. The main goal of the assignment is for students to deduce the gene map of one hybrid {"}Frankenplasmid{"} using the LacZ phenotype of its transformants, PCR, and restriction mapping. Our protocol results in a number of possible outcomes, meaning that students are mapping truly unknown plasmids. The open-ended nature of this assignment results in an effective module that teaches recombinant DNA procedures while engaging students with its investigative approach, increasing complexity, and puzzle-like quality. Moreover, the modular design of the activity allows it to be adapted to a more limited schedule, introductory courses, or more advanced courses.",
keywords = "PCR, Plasmids, Recombinant DNA, Restriction mapping",
author = "Dean, {Derek M.} and Wilder, {Jason A}",
year = "2011",
month = "9",
doi = "10.1002/bmb.20531",
language = "English (US)",
volume = "39",
pages = "376--383",
journal = "Biochemistry and Molecular Biology Education",
issn = "1470-8175",
publisher = "John Wiley and Sons Inc.",
number = "5",

}

TY - JOUR

T1 - The "Frankenplasmid" lab

T2 - An investigative exercise for teaching recombinant DNA methods

AU - Dean, Derek M.

AU - Wilder, Jason A

PY - 2011/9

Y1 - 2011/9

N2 - We describe an investigative laboratory module designed to give college undergraduates strong practical and theoretical experience with recombinant DNA methods within 3 weeks. After deducing restriction enzyme maps for two different plasmids, students ligate the plasmids together in the same reaction, transform E. coli with this mixture of ligated DNA, and plate the cells on media that specifically select for hybrid plasmids. The main goal of the assignment is for students to deduce the gene map of one hybrid "Frankenplasmid" using the LacZ phenotype of its transformants, PCR, and restriction mapping. Our protocol results in a number of possible outcomes, meaning that students are mapping truly unknown plasmids. The open-ended nature of this assignment results in an effective module that teaches recombinant DNA procedures while engaging students with its investigative approach, increasing complexity, and puzzle-like quality. Moreover, the modular design of the activity allows it to be adapted to a more limited schedule, introductory courses, or more advanced courses.

AB - We describe an investigative laboratory module designed to give college undergraduates strong practical and theoretical experience with recombinant DNA methods within 3 weeks. After deducing restriction enzyme maps for two different plasmids, students ligate the plasmids together in the same reaction, transform E. coli with this mixture of ligated DNA, and plate the cells on media that specifically select for hybrid plasmids. The main goal of the assignment is for students to deduce the gene map of one hybrid "Frankenplasmid" using the LacZ phenotype of its transformants, PCR, and restriction mapping. Our protocol results in a number of possible outcomes, meaning that students are mapping truly unknown plasmids. The open-ended nature of this assignment results in an effective module that teaches recombinant DNA procedures while engaging students with its investigative approach, increasing complexity, and puzzle-like quality. Moreover, the modular design of the activity allows it to be adapted to a more limited schedule, introductory courses, or more advanced courses.

KW - PCR

KW - Plasmids

KW - Recombinant DNA

KW - Restriction mapping

UR - http://www.scopus.com/inward/record.url?scp=80053116509&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80053116509&partnerID=8YFLogxK

U2 - 10.1002/bmb.20531

DO - 10.1002/bmb.20531

M3 - Article

C2 - 21948510

AN - SCOPUS:80053116509

VL - 39

SP - 376

EP - 383

JO - Biochemistry and Molecular Biology Education

JF - Biochemistry and Molecular Biology Education

SN - 1470-8175

IS - 5

ER -