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The Plant Health Instructor

Volume: 24
Year: 2024
Article Type: Lab Exercises
​​iTAG: Interactive Laboratory Exercises to Explore Genotype and Phenotype Using Oregon Wolfe Barley​

Appendix​​

​Roger P. Wise​,1,2,3 Gregory Fuerst,1Nick Peters,2 Nancy Boury,2 Laurie McGhee,4 Melissa Greene,5 Sarah Michaelson,6 Julie Gonzalez,7 Nick Hayes,8 Ron Schuck,9 Lance Maffin,10 Garrett Hall,11 Taylor Hubbard,12 and Ehren Whigham13​​

1 U.S. Department of Agriculture-Agricultural Research Service, Corn Insects and Crop Genetics Research Unit, Iowa State University, Ames, IA 50011, USA

2 Department of Plant Pathology, Entomology, and Microbiology, Iowa State University, Ames, IA 50011, USA

3 Correspondence to Roger.Wise@usda.gov

4 Colfax-Mingo Community High School, 204 N League Rd, Colfax, IA 50054, USA

5 Albia Community School District, 701 Washington Ave E, Albia, IA 52531, USA

6 Lake Forest Academy, 1500 W Kennedy Rd, Lake Forest, IL 60045, USA

7 Des Moines Area Community College, Des Moines, IA 50236, USA

8 Cedar Rapids Kennedy High School, 4545 Wenig Rd NE, Cedar Rapids, IA 52402, USA

9 (Retired) Ames Community High School, 1925 Ames High Dr, Ames, IA 50010, USA

10 Bondurant-Farrar Community High School, 1000 Grant St N, Bondurant, IA 50035, USA

11 Burr and Burton Academy, 57 Seminary Ave, Manchester, VT 05254, USA

12 Ankeny Community High School, 1155 SW Cherry St, Ankeny, IA 50023, USA

13 Creighton University, 2500 California Plaza, Omaha, NE 68178, USA​

Date Accepted: 21 Jan 2024
Date Published: 09 May 2024

Keywords: genotype, phenotype, Oregon Wolfe barley, epistasis, domestication, Genetics, Disease Resistance, homoeotic mutations





​Table 12. Equipment and materials supplied by instructor
Item​​​ When Needed:
20 x 13 cm (5-inch) pots​​​​​Growing plants​

Seed tray with 20 cells or 20 small pots
20-40 plant tags
Soil
Masking Tape
Fertilizer (20-20-20)
20 dowel rods or stakes
Twist ties or plant wire
Source of light
Gloves​​​​Throughout ​module
​ ​ ​ ​ ​
Goggles
10 fine point permanent markers
Refrigerator / Freezer
Ice (crushed is best)
Microcentrifuge tube rack
10-20 glass microscope slides​​​​DNA extraction
​ ​ ​ ​ ​
10-20 razor blades
10-20 glass stirring rods
70% or 90% rubbing alcohol
10-20 Styrofoam or plastic cups
Water bath
Balance or scale (more than one is helpful)DNA extraction & electr​ophoresis
Distilled water​​​​​Electrophoresis​
1 x 500 ml Erlenmeyer flask
1 x graduated cylinder (100ml)
Oven mitt or hot glove
Parafilm or wax paper
Lab tape (masking tape will work)
Microwave
Food coloring​Micropipette practice
Analytical balance
Weigh boats

 

Table 13. Core Equipment and Materials Provided in Instructor Travel Kits​
Oregon Wolfe barley seeds (20 plants)
All tubes (PCR tubes, 1.5 ml tubes, 2.0 ml tubes)
All solutions
Micropipettes and tips (4 sizes)
Thermal cycler
Vortex
Centrifuge
2 – Electrophoresis chambers & combs
1 – Electrophoresis power supply
Transilluminator

 

Table 14. iTAG su​pply list per 20 plant set ​ ​ ​ ​
Equipment Kap Vrs1 Practice Total
2.0 ml microcentrifuge tubes40  40
1.5 ml microcentrifuge tubes121240163
Small micropipette tips (10 µl)6813060258 (3 boxes)
Medium micropipette tips (200 µl)20442084 (1 box)
Large micropipette tips (101-1000 µl)149220171 (2 boxes)

Regular tube protocol:

PCR tubes

2040 60
PCR beads protocol: GE Healthcare: Illustra™ PureTaq™ Ready-To-Go™ PCR Beads (Store @ room temp)2020 40

PCR beads protocol:

PCR tubes

 20 20
Reagents ​ ​ ​ ​
2X CTAB buffer (Store @ room temp)19.6 ml  19.6 ml
20% SDS (Store @ room temp)2.1 ml  2.1 ml
5M potassium acetate (Store @ -20˚C)8.3 ml  8.3 ml
70% ethanol (Store @ -20˚C)10.1 ml  10.1 ml
Absolute isopropanol (Store @ -20˚C)11 ml  11 ml
TE buffer (Store @ room temp)4.1 ml  4.1 ml
2-Mercaptoethanol (Store @ room temp)405 µl  405 µl

 

OverviewExperiment 1Experiment 2
iTAG Instructor’s Planning Resources
AppendixGlossary​​​