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Team:Grinnell/Notebook
From 2011.igem.org
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===Week 2=== | ===Week 2=== | ||
| - | <html><ul style="list-style:none; float:right;"><li><img src="https://static.igem.org/mediawiki/2011/3/32/20110606_PCRproduct.jpg" alt="DNA gel from 20110606" height= | + | <html><ul style="list-style:none; float:right;"><li><img src="https://static.igem.org/mediawiki/2011/3/32/20110606_PCRproduct.jpg" alt="DNA gel from 20110606" height=200px/></li><li>PCR products on DNA gel. <br/>Lane 1: ladder; Lane 2: <i>rsaA</i> from<br/>liquid culture <i>Caulobacter</i>; Lane 3:<br/><i>rsaA</i> from plate culture <i>Caulobacter</i>;<br/>Lane 4: <i>esp</i> from <i>S. epidermidis</i>.</li></ul></html> |
Performed PCR on esp and rsaA genes ([[Team:Grinnell/Notebook/Protocols#cPCR|protocol]]) and ran results on gel ([[Team:Grinnell/Notebook/Protocols#dGel|protocol]]). Results (shown to the right) showed successful amplification. | Performed PCR on esp and rsaA genes ([[Team:Grinnell/Notebook/Protocols#cPCR|protocol]]) and ran results on gel ([[Team:Grinnell/Notebook/Protocols#dGel|protocol]]). Results (shown to the right) showed successful amplification. | ||
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Inoculated liquid cultures with colonies from transformations. | Inoculated liquid cultures with colonies from transformations. | ||
| - | Performed digestion of results from transformation. Results shown to the right. | + | Performed digestion of results from transformation and ran on gel. Results shown to the right. |
| - | <html><ul style="list-style:none; float:right"><li><img src="https://static.igem.org/mediawiki/2011/a/a5/20110610PlamidGel.jpg" alt="Plasmid result gel for 20110610" height= | + | <html><ul style="list-style:none; float:right"><li><img src="https://static.igem.org/mediawiki/2011/a/a5/20110610PlamidGel.jpg" alt="Plasmid result gel for 20110610" height=200px/></li><li>Gel results for transformation of ligations.<br/>Lane 1: ladder; Lane 2: digested plasmid<br/>with <i>rsaA</i>; Lane 3: digested plasmid with<br/><i>rsaA</i> and <i>esp</i>; Lanes 4-8: digested<br/>plasmids from various colonies with <i>esp</i>.</li></ul></html> |
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| + | Transformed BBa_K081005, a BioBrick part containing a gene for constitutive promotor and RBS, into ''E. coli'' Top10. | ||
Revision as of 21:53, 10 June 2011
Notebook
Week 1
PCR primers designed according to Biobrick specifications and ordered for rsaA C-terminal and WT esp. Because of the discrepancy in GC content between Staphylococcus epidermidis and Caulobacter, we also optimized the esp sequence for Caulobacter for later synthesis with a higher resulting GC content.
Prepared competent cells of E. coli Top10. (Protocol)
Streaked plates of Caulobacter, Staphylococcus aureus, and S. epidermidis.
Template DNA for PCR prepared from colonies using [http://www.bioventures.com/products.php?cid=10005| GeneReleaser]. (Protocol)
Week 2
- PCR products on DNA gel.
Lane 1: ladder; Lane 2: rsaA from
liquid culture Caulobacter; Lane 3:
rsaA from plate culture Caulobacter;
Lane 4: esp from S. epidermidis.
Performed a transformation experiment (protocol) to test the efficiency of our competent cells make in Week 1. Results were successful with about a 0.7 x 10^7 CFU/μg of pUC19.
Purified PCR product (protocol) and digested esp with EcoRI, SpeI, and PstI; rsaA with EcoRI, XbaI, and PstI; and pSB1C3 with EcoRI and PstI. We then ligated esp, rsaA, and esp and rsaA to pSB1C3 and transformed into E. coli Top10. (Protocol)
Inoculated liquid cultures with colonies from transformations.
Performed digestion of results from transformation and ran on gel. Results shown to the right.
- Gel results for transformation of ligations.
Lane 1: ladder; Lane 2: digested plasmid
with rsaA; Lane 3: digested plasmid with
rsaA and esp; Lanes 4-8: digested
plasmids from various colonies with esp.
Transformed BBa_K081005, a BioBrick part containing a gene for constitutive promotor and RBS, into E. coli Top10.
