Week 14: 9/26 - 10/2

9/26

9/27

9/28

9/29

Toggle 2 Gel Construct using Phire Polymerase

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

9/30

10/1

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: