IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2014/05/28

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Date: 5/28/14 People in lab: Emily Puleo, Caleb Treccazi

Title: PCR of hmp to add prefix and suffix Start Time: 10:35 am Purpose: Amplify hmp with iGEM prefix and suffix Protocol: LTM ed. 2 PCR Exceptions: 7.5 uL template E Coli DNA, 2.5 uL primers Products:

Sample Label Description Source Label Quantity
PCR hmp 5/28 A hmp with prefix and suffix at 55 C annealing temp --- 1
PCR hmp 5/28 B hmp with prefix and suffix at 55 C annealing tmep --- 1




Results: Notes: Accidentally put products in freezer during GE before TOPO cloning - CTZ Stop Time: 2:30 pm Next: Gel Electrophoresis to check products

Date: 5/28/14 People in lab: Emily Puleo, Caleb Treccazi

Title: Gel Electrophoresis of PCR hmp 5/28 A and B Start Time: 2:30 pm Purpose: Check for product Protocol: LTM Ed. 2 Gels Exceptions: Products:

Well 1 2 3 4 5 6 7 8
- - Ladder - PCR hmp 5/28 A - PCR hmp 5/28 B -

Products: Results: Notes: Used 90 mA for ~35 min Stop Time: 3:40 pm Next: TOPO



- Ladder - PCR hmp 5/28 A - PCR hmp 5/28 B - -

Date: 5/28/14 People in lab: Emily Puleo, Caleb Treccazi

Title: TOPO Cloning PCR hmp products from 5/28 and transform Start Time: 3:40 pm Purpose: Make more products Protocol: Invitrogent TOPO cloning kit manual and LTM Ed. 2 Chemical Transformations Exceptions: Products:

Sample Label Description Source Label Quantity
hmp TOPO 5/28 A1-1 Plated transformed cells on xgal amp plate 20 uL PCR hmp 5/28 A 1
hmp TOPO 5/28 B1-1 Plated transformed cells on xgal amp plate 20 uL PCR hmp 5/28 B 1
hmp TOPO 5/28 A1-2 Plated transformed cells on xgal amp plate 200 uL PCR hmp 5/28 A 1
hmp TOPO 5/28 B1-2 Plated transformed cells on xgal amp plate 200 uL PCR hmp 5/28 B 1
hmp TOPO 5/28 A2-1 Plated transformed cells on xgal amp plate 20 uL PCR hmp 5/28 A 1
hmp TOPO 5/28 B2-1 Plated transformed cells on xgal amp plate 20 uL PCR hmp 5/28 B 1
hmp TOPO 5/28 A2-2 Plated transformed cells on xgal amp plate 200 uL PCR hmp 5/28 A 1
hmp TOPO 5/28 B2-2 Plated transformed cells on xgal amp plate 200 uL PCR hmp 5/28 B 1

Results: Notes: Sample PCR hmp 5/28 A1 was melted in the hot water bath and the sample was vaporized; 15 uL 1000x xgal on amp plates for second batch; different ages of plates used for each batch Stop Time: 7:15 pm Next: Check growth and innoculate in LB+amp












































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