IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2013/08/08

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Date: 08/08/13 People in lab: Blythe Ferriere

Title: Blue/white screening for norV TOPO-TA cloned colonies

Start Time: 4:10PM

Purpose: Plate screened for blue white colonies white colonies should have plasmid.

Protocol: Colonies from plate CT1 8/7/13 were inoculated into LB broth + ampicillin tubes

Products:

Sample Label Description Source Label Quantity
8/8/13 I 1 Isolation and inoculation of white colony 8/7/13 CT1 8
8/8/13 I 9 Isolation and inoculation of light blue colony 8/7/13 CT1 1
8/8/13 I 10 Isolation and inoculation of blue colony 8/7/13 CT1 3

Notes: Ampicillin re-done in I1-I8

Stop Time: 5:41 PM

Next: Pick tubes with growth from white colonies for further analysis for check for norV gene


Date: 08/08/13 People in lab: Emily Puleo

Title: Gel electrophoresis of 8/7/13 PCR samples

Start Time: 4:37 PM

Purpose: To check for successful PCR reaction.

Protocol: LTM ed. 2 pg. 44

Well 1 2 3 4 5 6 7 8
Sample Ladder 8/7/13 A norCB 8/7/13 A nosZ 1 8/7/13 A nosZ 2 8/7/13 A hmp 1 8/7/13 A hmp 2

Results: Product from hmp only. PICTURE

Notes: Used 70 uL sample+LoadingDye per well

Stop Time: 5:20PM

Date: 08/08/13 People in lab: Emily Puleo

Title: TOPO-TA cloning of 8/7/13 PCR samples

Start Time: 5:05 PM

Purpose: To put genes into vectors and transform into bacteria to amplify.

Protocol: Invitrogen TOPO-TA cloning manual

Products:

Sample Label Description Source Label Quantity
8/8/13 hmp L1 hmp in TOPO-TA cloning vector 8/7/13 hmp A1 1

Notes: Samples were 1 day old only one was completed because only one ampicillin plate was ready

Stop Time: 6:15 PM

Date: 08/08/13 People in lab: Emily Puleo

Title: Transformation of 8/8/13 hmp L1

Start Time: 6:10 PM

Purpose: To put the hmp-TOPO vector into DH5alpha cells for growth and selection.

Protocol: LTM ed. 2 pg 41

Products:

Sample Label Description Source Label Quantity
8/8/13 hmp T hmp in TOPO-TA cloning vector in DH5alpha plated on LB+amp 8/8/13 hmp L1 1

Notes:

Stop Time: 9:30 PM


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