From 2009.igem.org
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| - | Descriptive Title of What You're Doing
| + | NotI restriction digest of isolated LuxOD47A on psB1AC3 constructs. |
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| - | WIKI CODING HERE
| + | Purpose: Even though the colony PCR from 2 days ago had negative control contamination, we’re going to run a NotI restriction digest anyway. If we can excise a gene of the appropriate size, it is likely that the sample in question actually contains LuxOD47A on psB1AC3, as opposed to mere master mix contamination. |
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| | + | Protocol: NotI restriction enzymes were used with REact I buffer, in accordance with the restriction digest procedure outlined on the protocol page. |
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| | + | Results: As shown below, the 4th set of digested genes displayed the set of bands that we expected: a band around 1400 bp (accounting for LuxOD47A) and a band around 3000 bp (accounting for psB1AC3). We will use this sample in our later constructions. |
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Revision as of 23:01, 20 August 2009
University of Calgary
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CAROL
Descriptive Title of What You’re Doing
WIKI CODING HERE
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CHINMOYEE
CLASS
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EMILY
Descriptive Title of What You're Doing
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FAHD
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IMAN
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JAMIE
Descriptive Title of What You're Doing
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JEREMY
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KATIE
Back to the Lab after Completing Initial Version of Quorum Sensing
Quorum sensing bacteria are complete and I made bacteria with RFP and placed them beside other bacteria for display. I will probably return to work on this later, but it is now more important to finish the lab. I learned that the order in which you add materials during restriction digest is important so using the function (initially from cleaning script) created for storing the names of items in an inventory will be important to make sure that each time something is added, it is the correct item. I believe for the rest of the activity I may be able to use a modified version of my find item script.
However, I have yet to determine a new way to move things to the water bath, heating block etc. in a creative way and still keep avatars involved.
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KEVIN
1. Plasmid Isolation of R0040, B0015, and J13002
R0040 (promoter), B0015 (terminator), and J13002 (RBS+Promoter) were isolated in order to verify the presence of them with Restriction digest. These parts are needed for general construction of our circuits.
2. Restriction digest of R0040, B0015, and J13002
To verify if B0015, R0040, and J13002 are present within the plasmid/cell, restriction digest was performed using NotI enzyme.
The band in R0040 C2 #1 is the only one that did not match the expected size of each. Only one colony is needed to move on.
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MANDY
Descriptive Title of What You're Doing
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PATRICK
Descriptive Title of What You're Doing
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PRIMA
Descriptive Title of What You're Doing
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STEFAN
Descriptive Title of What You're Doing
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VICKI
NotI restriction digest of isolated LuxOD47A on psB1AC3 constructs.
Purpose: Even though the colony PCR from 2 days ago had negative control contamination, we’re going to run a NotI restriction digest anyway. If we can excise a gene of the appropriate size, it is likely that the sample in question actually contains LuxOD47A on psB1AC3, as opposed to mere master mix contamination.
Protocol: NotI restriction enzymes were used with REact I buffer, in accordance with the restriction digest procedure outlined on the protocol page.
Results: As shown below, the 4th set of digested genes displayed the set of bands that we expected: a band around 1400 bp (accounting for LuxOD47A) and a band around 3000 bp (accounting for psB1AC3). We will use this sample in our later constructions.
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