IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2014/03/11

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Date: 03/11/14 People in lab: Levi Palmer

Title: Gel electrophoresis of 3/10 Digest

Start Time: 11:00 AM

Purpose: To check for presence of hmp gene in digest and TOPO vector

Protocol: LTM ed. 2

Results: No hmp present, no bands present, no picture taken

Next: Retry hmp PCR and TOPO cloning

Date: 03/11/14 People in lab: Levi Palmer

Title: Making chemically competent cells

Purpose: To make chemical competent cells to use in lab.

Protocol: OpenWetWare: Making Chemically Competent Cells (Inoue) Exceptions: 1. O.D. <0.1 of SOB grown culture 2. Adjusted to volume of 250mL 3. 1/2 volume of final resuspension

Products:

Sample Label Description Source Label
Chemical Competent 3/11/14 Chemically competent DH5alpha, in -80C DH5alpha stock

Notes: Test for competence was successful.

Date: 03/11/14 People in lab: Levi Palmer

Title: PCR of norCB, nosZ, hmp, norV from Genome Prep and E. coli colony PCR, then gel electrophoresis to confirm

Purpose: To amplify the genes and check for successful PCR, also a check for successful genome prep.

Protocol: LTM ed. 2 pg 48, 44-46


Results: No bands visible for norV, norCB, nosZ. Strong band visible for hmp

Well 1 2 3 4 5 6 7 8
Sample norV hmp norCB Ladder RGTT LMPG

Notes: The wrong organism was used for norV, that is why there was no product. norV is an E. coli gene. RGTT and LMPG are mini-project parts.

Stop Time: 8:00PM

Next: Retry PCR with Dr. Westenberg's primers for bacterial DNA (S16 Ribosome subunit)


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