The document outlines the equipment requirements for two practical microbiology classes involving DNA extraction and PCR. For a DNA extraction class with 35 students, it lists the reagents needed for each student to extract DNA from sediment samples using a commercial kit. This includes gloves, safety glasses, balances, centrifuges, pipettes, tip boxes, reagent aliquots, and storage tubes. For a subsequent PCR class, it details the equipment needed for students to perform PCR on their extracted DNA and run a gel, including PCR machines, electrophoresis kits, centrifuges, reagents for running PCR and gel, and supplies for preparing the gel. Student reagents will be aliquoted into racks prior to the class.

1. Equipment Requirements for Microbial Ecology Practical Classes
(24/6/13)
28 students enrolled, class size cap of 35 – perhaps plan for 35 students to be
safe.
Prac 3: DNA Extraction (Aug 19, 2012):
Description:
Students will extract microbial genomic DNA from the sediment samples they collected in
Practical 1 using a commercial DNA extraction kit (MoBio Power soil DNA Isolation kit [$529
- $5.29 per extraction. Students will be given enough material for two extractions each]).
Required aliquots of each reagent will be dispensed prior to class so students each have
own stock of each reagent
Equipment required:
- Sediment sample from Prac 1
- Several boxes of rubber gloves of different sizes
- 35 x Safety glasses
- 10-20 x spatula (small – only 0.25 g of sediment collected)
- 10 x laboratory balance
- 50 x cake holders, cups/trays for weighing sediment in
- 20 x fine black textas
- 5 x small centrifuges (to fit 2 ml eppindorf tubes)
- MoBio vortex unit (JS can provide 2)
- 10 x vortexes
- 30 x 20-200ul pipettes
- 30 x 200-1000ul pipettes
- 15 x blue pipette tip boxes
- 15 x yellow pipette tip boxes
- 5 x tip disposal boxes
- 35 x plastic racks (1 per student) holding labelled aliquots from MoBIO DNA extraction kit
(for each student, in 2ml eppindorf tube):
120 ul C1
500 ul C2
400 ul C3
2 ml C4
1000 ul C5
200 ul C6

2. 190 ul of Qubit working solution, pre-mixed and in Qubit tube (Mix of ds DNA HS
assay kit reagents)
- For each student: 1 bead tube, 1 spin filter, 5 x 2ml (eppindorf tubes) collection tubes [All
from kit]
- 5 x Qubit specs
- Access to refrigerator
Post-prac requirements:
DNA samples (20 - 25 x 2ml eppindorf tubes) will need to be stored in freezer for use in
Prac. 5
Prac 5: PCR (Sept 2, 2012):
Description:
Students will perform a PCR on the DNA extracted in Prac 4 and subsequently run a gel to
visualise amplified DNA.
PCR cycle 1:34 cycle (To be programmed prior to prac – JS to provide cycling conditions)
Equipment required:
- DNA from Prac 3
- 2-4 PCR machines
- 15 electrophoresis kits/gel baths
- 3 gel illuminators
- 5 microcentrifuges that fit PCR tubes
- 35 x plastic racks (1 per student) holding 5 PCR tubes + eppindorfs containing enough to do
4 PCR reactions (sample, positive control, negative control), per reaction:
25 ul GoTaq Green Master mix
2 ul primer mix
10 ul DNA
13 ul MilliQ
 So each student will require eppendorf tubes containing:
100 ul GoTag Green Master mix
10 ul primer mix (27F, 1492R – JS to provide details)
100 ul MilliQ
15 ul positive control DNA (JS to provide DNA)
10 ul of Gel Red
Ladder (JS to provide details)
- 30 x 2-20 ul pipettes
- 30 x 20-200 ul pipettes

3. - 30 x 1 ml pipettes
- 30 x thin markers.. suitable for labelling PCR tubes
- 2-3 containers of agarose for gels
- 10-15 x 300 ml TAE for gels
-15 x 100 ml Schott bottles for mixing up gel
- 10 x 100 ml measuring cylinders for measuring TAE
- 5 x spatula big
- 4 x lab balance
- 3 x microwave
- 10 x 500 ml sterile schott bottles