Team:Stony Brook/Experiments

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Protocols & Experiments

Universal Lab Protocols

Agarose Gel Preparation

Add 50mL of 1X TAE buffer to a 500mL erlenmeyer flask
Add 0.5g of 1% agarose
Microwave for three 20 second intervals and cool slightly
Add 1ul Ethidium Bromide
Mix and pour into gel box. Add Comb
Cool for 30 minutes

YPD Media

For liquid media

Add 20g of Bacto peptone, 10g of yeast extract and 950mL of water to a flask
Autoclave on the liquid setting
Add 50mL of 40% glucose and mix

For solid media
- Add same reagents as liquid, plus 24g of Bacto agar
- Autoclave
- Stir on a magnetic stir plate and add 50ml of 40% glucose
- Pour into petri dishes

LB Broth

Add items to flask:

5g Tryptone
2.5g Yeast Extract
5g NaCl
H₂O filled to 500mL
Mix with magnetic stir bar on low heat, autoclave for liquids

Nanodrop

Select Nucleic Acid
Clean, drop 1ul water, measure
Blank
Measure with 1ul of substance
For PCRs: 260/280 ratio should be near 2

LB Agar

Add items to flask:

5g Tryptone
2.5g Yeast Extract
5g NaCl
7.5g Agar
H₂O filled to 500mL
Mix with magnetic stir bar on low heat, autoclave for liquids

NEB Monarch Nucleic Acid Purification Protocols

The following NEB kits were used and protocols can be found on their website

Monarch Plasmid Miniprep Kit
Monarch DNA Gel Extraction Kit
Monarch PCR & DNA Cleanup Kit

Transformation

Thaw competent E. coli cells on ice and pipette 50ul cells into a transformation tube
Add 1-5ul containing 1pg-100ng of plasmid DNA and flick tube 4-5 times
Place tube on ice for 30 minutes
Heat shock at 42°C for 30 seconds
Place on ice for 5 minutes
Add 950ul room temperature SOC to mixture
Incubate at 37°C for 60 minutes at 250rpm
Mix and perform several 10-fold serial dilutions in SOC
Spread 50-100ul onto a plate and incubate overnight at 37°C

Cancer Group Protocols

Vaccine Group Protocols

Inspiration