pGLO™ SDS-PAGE Extension

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pGLO SDS-PAGE Extension

Overview

Lab Preparation Checklist

Kit contains sufficient materials for 8 student workstations (2–4 students per workstation)

Dithiothreitol (DTT), 0.3 g
1
Precision Plus Protein Kaleidoscope standards, 50 µl  2
Bio-Safe Coomassie stain for proteins, 100 ml
1
10x Tris-glycine-SDS electrophoresis buffer, 1 L
1
Laemmli sample buffer, 30 ml
1
Curriculum, including Teacher's Guide and Student Manual
1

Required Accessories Not Included in Kit:

Amp (white) and amp/ara (green) colonies from plates from the pGLO bacterial transformation lab 4–8
Ready Gel® or Mini-PROTEAN® TGX precast polyacrylamide gels, 4–20%, 10-well each 4–8
Vertical gel electrophoresis chambers 1–4
Power supplies 1–4
Water bath or dry bath 1
Adjustable micropipets
   2–20 µl
   100–1,000 µl

4–8
4–8
Pipet tips
   2–200 µl, BR-35
   100–1,000 µl, BR-40
   0.5–200 ul, Prot/Elec

1 bag
1 bag
1 bag
Screwcap microcentrifuge tubes, 1.5 ml 500

Recommended (Optional) Accessories:

GelAir drying system
GelAir assembly table
GelAir drying frames with clamps
GelAir cellophane
Molecular Imager® GelDoc XR system
pGLO Kit SDS-PAGE Extension Flowchart
pGLO SDS-PAGE Extension Flowchart
Key Kit Features
  • Perform real biotechnology workflows
  • Identify proteins by conformation and size
  • Link gene induction to protein expression to protein identification
  • Understand chromophores and the basis of protein fluorescence

The pGLO™ Kit SDS-PAGE Extension is designed to be performed following the pGLO Bacterial Transformation Kit. The extension is designed to identify and visualize green fluorescent protein (GFP) from the proteome of transformed E. coli, which also contains thousands of other proteins. Because only the cloned GFP glows, students can use this phenotypic trait to quickly identify the GFP within the complex mix of proteins.

Features and Benefits

With this kit, students are able to:

  • Perform real biotechnology workflows
  • Identify proteins by conformation and size
  • Link gene induction to protein expression to protein identification
  • Understand chromophores and the basis of protein fluorescence
  • Extend the pGLO Bacterial Transformation Kit

How It Works

In the first part of the exercise, students use the pGLO Bacterial Transformation Kit to transfer pGLO plasmid encoding GFP into E. coli, a common prokaryotic organism used for DNA propagation and protein expression. Colonies of E. coli are qualitatively examined for fluorescence to determine whether the pGLO gene is being expressed. In the second part of the laboratory activity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is employed to separate the entire repertoire of proteins expressed in E. coli, including the foreign GFP responsible for transferring the fluorescence trait.

More Information

  • Three-session laboratory activity, 45 min per session
  • Provides sufficient materials for eight student workstations, up to four students per workstation.
Kit contains sufficient materials for 8 student workstations (2–4 students per workstation)  
Laemmli sample buffer, 30 ml 1
Precision Plus Protein™ Kaleidoscope™ prestained standards, 50 µl 2
Dithiothreitol (DTT), 0.3 g 1
10x Tris-glycine-SDS electrophoresis buffer, 1 L 1
Bio-Safe™ Coomassie stain for proteins, 100 ml 1
Curriculum, including Teacher's Guide and Student Manual 1
pGLO Kit SDS-PAGE Extension

166-0013EDU
Protein analysis classroom study kit, includes DTT, protein standards, Bio-Safe Coomassie stain, 10x TGS, Laemmli sample buffer, curriculum, for 32 students; education use only

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M1660023 Instruction Manual, Protein Electrophoresis of GFP: A pGLO Bacterial Transformation Kit Extension, Biotechnology Explorer, Rev D Please login to download this document