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id6-17
Propagating MAV1212 and MAV1212-Hef1a-EBFP2 (Begun 6-17)

Stock Concentrations:

EBFP2: 1118 ng/µL

MAV: unknown.

1856: Diluted DNA samples for both x100 before transformation

4 Plates, LO HI for both.

2144: Culture plates incubated overnight at 30°C for 16-18 hours.

 

 

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id6-18
Propagating MAV1212 and MAV1212-Hef1a-EBFP2 (Begun 6-17)
1426: Checked cell plates. EBFP2 produced colonies. MAV1212 overgrown.
MAV1212 Streaking (Start 6-18)Propagating EBFP2: Pick Colonies (Start 6-17)
2044: Picked single colonies from each plate and streaked over xgal plates.
Let on the incubator overnight at 30°C for 16-18 hours.		2040: Picked one colony each from the HI and LO plates and immersed in LB Media.
Left on the shaker incubator overnight.

 

 

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id6-19
MAV1212 Streaking (Start 6-18) OVER
1254: Disaster. No colonies.
Propagating MAV1212 and MAV1212-Hef1a-EBFP2 (AgainStart 6-19)

Concentrations:

EBFP2: 12 ng/µL

MAV: 4 ng/µL

 

4 Plates, LO HI for both.

1715: Culture plates inseerted into 30°C incubator.

 

 

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id6-20
Propagating MAV1212 and MAV1212-Hef1a-EBFP2 (AgainStart 6-19) OVER

1105: Low transformation rate. Suspicions fall on faulty plates. Plating to be redone later with existing samples and streaking.

 

 

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id6-23

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id6-23
Propagating MAV1212: Colony Picking MAV1212(Start 6-17)Propagation MAV1212 (Start 6-23)

Attempted to isolate monoclonal colony from MAV1212 lawn.

Inserted into Amp+ LB medium.

Left on incubator shaker at 2114.

Re-plated transformation suspension on Amp+ plates.

Streaked colonies from lawn.

Left on 30°C incubator at 2120.

 

 

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id6-24

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id6-24
Propagating MAV1212: Colony Picking MAV1212(Start 6-17) OVERPropagation MAV1212 (Start 6-23) OVER

Medium Clear.

Growth failed4.

Current MAV1212 vectors abandoned due to overactive ccdB gene killing off cells.

Awaiting arrival of new vectors with better survivability.

Propagation MAV1212-EBFP2 (Start 6-24)

Transformed onto Amp+ plates.

Left on 30°C incubator at 2135.

 

 

14-6-25

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id6-25

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id6-25
Propagation MAV1212-EBFP2 (Start 6-24)
Success. (Like we need more EBFP2)