"
Team:Penn State/Notebook
From 2014.igem.org
(Difference between revisions)
| Line 102: | Line 102: | ||
<tr> | <tr> | ||
<td><center><b>Wednesday, May 21, 2014</b></center></td> | <td><center><b>Wednesday, May 21, 2014</b></center></td> | ||
| - | <td><b>Emily's first experience with cloning!</b> Ashlee led Emily through several practice experiments from designs made earlier in the year: making a gel, loading samples, gel purifying DNA</td> | + | <td><b>Emily's first experience with cloning!</b> Ashlee led Emily through several practice experiments from designs made earlier in the year: making a gel, loading samples, gel purifying DNA.</td> |
<td>...................</td> | <td>...................</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><center><b>Thursday, May 22, 2014</b></center></td> | <td><center><b>Thursday, May 22, 2014</b></center></td> | ||
| - | <td>Ashlee and Emily performed a transformation</td> | + | <td>Ashlee and Emily performed a transformation.</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><center><b>Friday, May 23, 2014</b></center></td> | <td><center><b>Friday, May 23, 2014</b></center></td> | ||
| - | <td>We picked several colonies for overnight growth to do more cloning tomorrow</td> | + | <td>We picked several colonies for overnight growth to do more cloning tomorrow.</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><center><b>Saturday, May 24, 2014</b></center></td> | <td><center><b>Saturday, May 24, 2014</b></center></td> | ||
| - | <td>Memorial Day Weekend? How about lab cloning weekend! Ashlee conducted plasmid preparation and digestion</td> | + | <td>Memorial Day Weekend? How about lab cloning weekend! Ashlee conducted plasmid preparation and digestion.</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td><center><b> | + | <td><center><b>Monday, May 26, 2014</b></center></td> |
| - | <td>Due to a string of failed clonings with the broadhost vector pSEVA251 and the large inserts, new designs are evaluated! Instead of creating a plasmid with the HMF pathway (7.5 kb) and dCas9 system (5.5 kb), we shall add the HMF pathway and dCas9 to the <i>P. putida</i> genome using homologous recombination. | + | <td>Due to a string of failed clonings with the broadhost vector pSEVA251 and the large inserts, new designs are evaluated! Instead of creating a plasmid with the HMF pathway (7.5 kb) and dCas9 system (5.5 kb), we shall add the HMF pathway and dCas9 to the <i>P. putida</i> genome using homologous recombination. </td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td><center><b> | + | <td><center><b>Tuesday, May 27, 2014</b></center></td> |
| - | <td>Inoculated LB broth with ampicillin and dCas9 plasmid from cryogenic storage; inoculated Lb broth with chloramphenicol and FTV vector from Ashlee's past experiment; streaked the HMF vector on a kanamycin plate</td> | + | <td>Inoculated LB broth with ampicillin and dCas9 plasmid from cryogenic storage; inoculated Lb broth with chloramphenicol and FTV vector from Ashlee's past experiment; streaked the HMF vector on a kanamycin plate.</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td><center><b> | + | <td><center><b>Wednesday, May 28, 2014</b></center></td> |
| - | <td>Emily and Ashlee made cryogenic storage of the dCas9 plasmid; plasmid prepared the FTV and dCas9 vectors; digested FTV vector; inoculated LB broth with a colony from the HMF plate for overnight growth and plasmid preparation tomorrow</td> | + | <td>Emily and Ashlee made cryogenic storage of the dCas9 plasmid; plasmid prepared the FTV and dCas9 vectors; digested FTV vector; inoculated LB broth with a colony from the HMF plate for overnight growth and plasmid preparation tomorrow.</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
<tr> | <tr> | ||
<td><center><b>Thursday, May 29, 2014</b></center></td> | <td><center><b>Thursday, May 29, 2014</b></center></td> | ||
| - | <td> | + | <td>Prepared plasmid containing the HMF pathway; inoculated LB broth with Lambda Red Recombinase plasmid from cryogenic storage. Ashlee, Emily, and graduate student Iman Farasat ordered primers for three plasmids that will be constructed via Gibson Chew-Back and Annealing Assembly, two of which will be inserted into the genome by homologous recombination.</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><center><b>Friday, May 30, 2014</b></center></td> | <td><center><b>Friday, May 30, 2014</b></center></td> | ||
| - | <td> | + | <td>Ashlee and Emily made cryogenic storage of the Lambda Red Recombinase plasmid.</td> |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><center><b>Monday, June 2, 2014</b></center></td> | <td><center><b>Monday, June 2, 2014</b></center></td> | ||
| - | <td></td> | + | <td>Constructed dCas9 gene cassette and plasmid backbone with replication origin ColE1 via PCR Rescue. Gel purified dCas9 and ColE1 cassettes.</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><center><b>Tuesday, June 3, 2014</b></center></td> | <td><center><b>Tuesday, June 3, 2014</b></center></td> | ||
| - | <td></td> | + | <td>Conducted Colony PCR using <i>P. putida</i> KT2440 strain as DNA template to construct two ~1 kb genome overlaps. Plasmid prepared the Lambda Red Recombinase plasmid, DH10B-PKD46, FTV-ptac-LacI-CmR plasmid, and NoHP_15A_Plmra_CmR plasmid containing RFP with a strong, unique promoter. Stock of NoHP_15A_Pkmra_CmR and FTV_ptac_LacI_CmR for cryogenic storage was also made. Lambda Red Recombinase cassette was amplified using PCR Rescue and gel purified.</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Revision as of 17:51, 20 June 2014
WELCOME TO iGEM 2014!Your team has been approved and you are ready to start the iGEM season!
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Penn State iGEM 2014 Notebook PageHere you will find weekly summaries of our wet laboratory progress, team updates, and accomplishments outside the laboratory. Below [link this] is our detailed, day-to-day progress laboratory notebook. Weekly Summarieswill create links to each week, but will just list down the page Week 1, Week 2, etc etc Laboratory Notebook
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
