Team:Grinnell/Notebook

From 2011.igem.org

(Difference between revisions)
(→Week 2)
Line 14: Line 14:
===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=250px/></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>
<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=250px/></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 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.
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/&mu;g 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/&mu;g of pUC19.
-
Purified PCR product ([[Team:Grinnell/Notebook/Protocols#dClean|protocol]]) and [[Protocols#Restriction_Digest|digested]] ''esp'' with EcoRI, SpeI, and PstI; ''rsaA'' with EcoRI, XbaI, and PstI; and spB1C3 with EcoRI and PstI.  We then [[Protocols#Ligation|ligated]] ''esp'', ''rsaA'', and ''esp'' and ''rsaA'' to spB1C3 and transformed into ''E. coli'' Top10. ([[Team:Grinnell/Notebook/Protocols#Transformation|Protocol]])
+
Purified PCR product ([[Team:Grinnell/Notebook/Protocols#dClean|protocol]]) and [[Protocols#Restriction_Digest|digested]] ''esp'' with EcoRI, SpeI, and PstI; ''rsaA'' with EcoRI, XbaI, and PstI; and pSB1C3 with EcoRI and PstI.  We then [[Protocols#Ligation|ligated]] ''esp'', ''rsaA'', and ''esp'' and ''rsaA'' to pSB1C3 and transformed into ''E. coli'' Top10. ([[Team:Grinnell/Notebook/Protocols#Transformation|Protocol]])
Inoculated liquid cultures with colonies from transformations.
Inoculated liquid cultures with colonies from transformations.
-
<html><ul style="list-style:none; float:left"><li><img src="https://static.igem.org/mediawiki/2011/a/a5/20110610PlamidGel.jpg" alt="Plasmid result gel for 20110610" height=250px/></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>
+
Performed digestion of results from transformation.  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=250px/></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>

Revision as of 21:45, 10 June 2011

Grinnell Menubar

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

  • DNA gel from 20110606
  • 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 PCR on esp and rsaA genes (protocol) and ran results on gel (protocol). Results (shown to the right) showed successful amplification.

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. Results shown to the right.

  • Plasmid result gel for 20110610
  • 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.

Retrieved from "http://2011.igem.org/Team:Grinnell/Notebook"