Team:Evry/Notebook

   

Notebook

Here is our lab notebook. Follow all the wet lab experiments we did, day by day.

• Yeast surface-display
  • May
    • Example entry
    • Week 2
    • Week 3
    • Week 4
• Yeast encapsulation
  • May
    • Week 1
    • Week 2
    • Week 3
    • Week 4
• Hypoxia bio-sensor
  • May
    • Week 1
    • Week 2
    • Week 3
    • Week 4
• MAIT cells stimulation
  • May
    • Week 1
    • Week 2
    • Week 3
    • Week 4
• Antigene prediction
  • May
    • Week 1
    • Week 2
    • Week 3
    • Week 4

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Friday, 12^th June 2015

Extraction of AgA1P from yeast genome with BSAI
AGA1P was extracted with BSAI overhangs for subsequent cloning from W303 and BY4000, according to Looke et al., PMC 2011.

1. Resuspend one yeast colony in 100 µl of 200 mM LiAc, 1% SDS solution and incubate at 70°C
2. Add 300 µl of 36% ethanol and vortex
3. Spin down DNA at 15 000 g for 3 minute
4. Wash pellet with 70% ethanol
5. Disolve pellet in 100 µl of water and spin down debris at 15 000 g for 15 seconds

PCR of AGA1P with primers AGA1P R1/F1 with Q5 polymerase using a gradient (57/60/63°C)

Q5 PCR Program

• step 1 : 98°C – 30 seconds
• step 2 : 98°C – 10 seconds
• step 3 : 57, 60 or 63°C
• step 4 : 72°C – 1 min (repeat steps 2-4 for 40 cycles)
• step 5 : 16°C - Hold

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Example notebook entry

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