IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2014/05/29

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Date: 5/29/14 People in lab: Emily Puleo, Caleb Treccazi Title: Checking plates from 5/28 Start Time: 11 am Purpose: Screen plates for correct colonies from 5/28 Protocol: Exceptions: Products:

Results: No growth Notes: Possible problemw with resistance or something wrong with xgal or a problem with freezing PCR product before TOPO Stop Time: 11:15 am Next: Try PCR again without accidental freezing and plate on both xgal amp and just amp plates to test resistances

Date: 5/29/14 People in lab: Emily Puleo, Caleb Treccazi

Title: PCR of hmp to add prefix and suffix Start Time: 11:20 am Purpose: Amplify hmp with the iGEM prefix and suffix Protocol: LTM Ed. 2 PCR Exceptions: 7.5 uL template, 2.5 uL primers Products:

Sample Label	Description	Source Label	Quantity
PCR hmp 5/29 A	hmp with iGEM prefix and suffix	---	1
PCR hmp 5/29 B	hmp with prefix and suffix	---	1

Results: Notes: Stop Time: 2 pm Next: Gel electrophoresis to check product

Date: 5/29/14 People in lab:Caleb Treccazi

Title: Gel Electrophoresis of PCR hmp 5/29 A and B Start Time: 2:05 pm Purpose: To check for successful products Protocol: LTM Ed. 2 Gels Exceptions: Products:

Well	1	2	3	4	5	6	7	8
-	-	Ladder	-	PCR hmp 5/29 A	-	PCR hmp 5/29 B	-

Products: Results: Notes: Used 90 mA for ~45 min Stop Time: 2:55 pm Next: TOPO

Date: 5/29/14 People in lab: Caleb Treccazi

Title: TOPO cloning of PCR hmp 5/29 A and B Start Time: 3:05 pm Purpose: To make copies of successful products Protocol: Invitrogen TOPO kit instruction manual Exceptions: Products:

Sample Label	Description	Source Label	Quantity
TOPO A 5/29	hmp product cloned	PCR hmp 5/29 A	1
TOPO B 5/29	hmp product cloned	PCR hmp 5/29 B	1

Results: Notes: Cloned for ~ 9 min Stop Time: 3:15 pm Next: Chemical Transformations

Date: 5/29/14 People in lab: Caleb Treccazi

Title: Chemical Transformation of TOPO A and B 5/29 Start Time: 3:20 pm Purpose: To grow and screen cells with hmp with the prefix and suffix Protocol: LTM Ed. 2 Exceptions: Products:

Sample Label	Description	Source Label	Quantity
CT 1 5/29	20 uL CT cells on amp xgal plate	TOPO A 5/29	1
CT 2 5/29	200 uL CT cells on amp xgal plate	TOPO A 5/29	1
CT 3 5/29	20 uL CT cells on amp xgal plate	TOPO B 5/29	1
CT 4 5/29	200 uL CT cells on amp xgal plate	TOPO B 5/29	1
CT 5 5/29	20 uL CT cells on amp plate	TOPO B 5/29	1
CT 6 5/29	200 uL CT cells on amp plate	TOPO B 5/29	1

Results: CT 5 was discarded as contaminated after lawn growth, plates with colonies were saved from CT 1 - CT 4 for screening Notes: Amp plates were unmarked or badly marked in the fridge but were assumed to be amp and usable. Stop Time: 6:10 pm Next: Save plates for isolations

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