Team:Stony Brook/Notebook/Cancer-W14

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Week 14: 9/26 - 10/2

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9/26

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9/27

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9/28

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9/29

Toggle 2 Gel Construct using Phire Polymerase

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PCR Ran of Toggle 2:

Phase	Temperature (°C)	Time (sec)
Initial Denaturation	98	60s
Denaturation	98	20s
Annealing	Ranged from 72-60 degrees for each tube. Did a different annealing temperature for each PCR.	10s
Extension	72	#
Final Extension	72	#

Ran 32 cycles

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9/30

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10/1

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10/2

Transformations

1. Thaw TSS cells on ice
2. Add 2ul of prepped plasmid. Pipette to mix
3. Sit for 30 minutes on ice
4. Incubate cells for 30s at 43°C
5. Incubate on ice for 2 minutes
6. Add 1ml of 50C
7. Incubate for 30 minutes at 37°C. Shake
8. Spread 100-300ul on plates
9. Grow overnight at 37°C
10. Save the rest as liquid cultures

Plates Used:

• Plate 1: pEPGAP
• Plate 2: pEPGAP/CRISBU
• Plate 3: pEPGAP-P
• Plate 4:
• Plate 5: pSBIA2/T1/T2
• Plate 6: pSBIA2
• Plate 7: pSBIA2-P
• Plate 8: