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Team:Grinnell/Notebook
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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=200px/></li><li>PCR products on DNA gel. | + | <html><ul style="list-style:none; float:right;"><li><a href="https://2011.igem.org/File:20110606_PCRproduct.jpg"><img src="https://static.igem.org/mediawiki/2011/3/32/20110606_PCRproduct.jpg" alt="DNA gel from 20110606" height=200px/></a></li><li>Figure 1: PCR products on DNA gel. Lane 1: ladder; Lane 2: <i>rsaA</i> from liquid culture <i>Caulobacter</i>; Lane 3: <i>rsaA</i> from plate culture <i>Caulobacter</i>; Lane 4: <i>esp</i> from <i>S. epidermidis</i>.</li><li><a href="https://2011.igem.org/File:20110610PlamidGel.jpg"><img src="https://static.igem.org/mediawiki/2011/a/a5/20110610PlamidGel.jpg" alt="Plasmid result gel for 20110610" height=200px/></a></li><li>Figure 2: Gel results for transformation of ligations. Lane 1: ladder; Lane 2: digested plasmid with <i>rsaA</i>; Lane 3: digested plasmid with <i>rsaA</i> and <i>esp</i>; Lanes 4-8: digested plasmids from various colonies with <i>esp</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 | + | 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 showed successful amplification (figure 1). |
Performed a transformation experiment ([[Team:Grinnell/Notebook/Protocols#Transformation|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. | Performed a transformation experiment ([[Team:Grinnell/Notebook/Protocols#Transformation|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. | ||
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Inoculated liquid cultures with colonies from transformations. | Inoculated liquid cultures with colonies from transformations. | ||
| - | Performed digestion of results from transformation and ran on gel | + | Performed digestion of results from transformation and ran on gel (figure 2). |
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Transformed BBa_K081005, a BioBrick part containing a gene for constitutive promotor and RBS, into ''E. coli'' Top10. | Transformed BBa_K081005, a BioBrick part containing a gene for constitutive promotor and RBS, into ''E. coli'' Top10. | ||
Revision as of 22:11, 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
- Figure 1: 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.
- Figure 2: 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.
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 (figure 2).
Transformed BBa_K081005, a BioBrick part containing a gene for constitutive promotor and RBS, into E. coli Top10.
