Jessica Karen Wong/Notebook/2007-8-6: Difference between revisions

Revision as of 14:05, 6 August 2007

Ran a gel of overnight pcr's
large gel:
- Row 1 : E0240 M1 ES (4-1), E0240 M2 ES (1-4), SP LAD SP E0240-ES-E2 (4-1)E0240-EX-E2 (1-4)
- Row 2 : F2620-3K3-1 (4-1), F2620-3K3-2 (1-4), SP LAD SP F2620-3K3-3 (4-1) E0240 M2-EX (1-4)

small gel: LAD SP E0240-E1-EX (4-1), E0240-EX-E1 (2-1), E0240-3X-M1 (2-1)
Overnighted F2620-3K3-2 colony 4, e0240-ES (M1 colony 1, M2 col 2, E2 col 1),E0240-EX (E2 col 2, E1 col 2)

Took out overnight liquid cultures and saw that I2057-EX-J116 was the only RFP culture that was red
Minipreped sealed I2055 EX, sealed I2055 SX, I2057-J116
Sent those 3 for sequencing with VF and VR
Made new I2057-EX-J100, I2057-EX-R0040, I2057-SX-R0040 from red colonies on each plate

Looked at the I2055-various RBS plates on the gel imager and saw very few fluorescent colonies
Realized that we forgot the restriction enzyme when ligating the RBS's in
Made an overnight of I2055-SX-B0031 which fluoresced
Streaked out a plate of I2055-SX-B0030 that had a small fluorescent area

Religated B0032-I2055-EX
Added .5ul of each S and X restriction enzyme to ligation mix of I2055-SX-B0032, 31, and 30
Transformed all 4 ligations into Top10 and plated

@@ Line 1: / Line 1: @@
+[[Image:8-6-large.jpg|thumb|left|Large Gel]]
 *Ran a gel of overnight pcr's
 *large gel:
 **Row 1 : E0240 M1 ES (4-1), E0240 M2 ES (1-4), SP LAD SP E0240-ES-E2 (4-1)E0240-EX-E2 (1-4)
 **Row 2 : F2620-3K3-1 (4-1), F2620-3K3-2 (1-4), SP LAD SP F2620-3K3-3 (4-1) E0240 M2-EX (1-4)
+[[Image:8-6-small.jpg|thumb|left|Small Gel]]
 *small gel: LAD SP E0240-E1-EX (4-1), E0240-EX-E1 (2-1), E0240-3X-M1 (2-1)
 *Overnighted F2620-3K3-2 colony 4, e0240-ES (M1 colony 1, M2 col 2, E2 col 1),E0240-EX (E2 col 2, E1 col 2)