IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2014/02/05

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Date: 02/5/14 People in lab: Kelsey Crossen

Title: PCR of nosZ, adding prefix/suffix

Start Time: 12:05 PM

Purpose: To amplify the nosZ gene and add the iGEM prefix/suffix

Protocol: LTM ed. 2 and Dr. Shannon's lab PCR protocol Exceptions: 1. Annealing temp: 50C, 2. Final primer conc. <0.1uM

Products:

Sample Label Description Source Label Quantity
nosZ 1 LTM protocol amplified nosZ P aeru 7//31 1
nosZ 2 Dr. Shannon lab protocol amplified nosZ P aeru 7/31 1

Notes: Dr Shannon's Protocol: 7.5 uL template, 3 uL DreamTaq buffer, 3 uL dNTPs, 2.5 uL F primer, 2.5 uL R primer, 10.5 uL milliQ, 1 uL DreamTaq. Hot start (95C) then add Taq.

Stop Time:

Next: Gel electrophoresis of PCR product

Date: 02/5/14 People in lab: Levi Palmer

Title: Gel electrophoresis of 2/5 PCR product

Start Time: 4:00 PM

Purpose: To check for successful PCR

Protocol: LTM ed. 2

Well 1 2 3 4 5 6 7 8
Sample Ladder nosZ1 nosZ2

Results: No product visible

Notes: Only 30sec elongation time, extend time and try again.

Stop Time: 8:00 PM


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