Team:UGA-Georgia/Notebook

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<!-- *** What falls between these lines is the Alert Box!  You can remove it from your pages once you have read and understood the alert *** -->
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= Notebook =
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== '''Methanococcus Lab''' ==
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<div id="template" style="text-align: center; font-weight: bold; font-size: large; color: #f6f6f6; padding: 5px;">
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This is a template page. READ THESE INSTRUCTIONS.
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<div id="instructions" style="text-align: center; font-weight: normal; font-size: small; color: #f6f6f6; padding: 5px;">
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You are provided with this team page template with which to start the iGEM season.  You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki.  You can find some examples <a href="https://2008.igem.org/Help:Template/Examples">HERE</a>.
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<div id="warning" style="text-align: center; font-weight: bold; font-size: small; color: #f6f6f6; padding: 5px;">
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You <strong>MUST</strong>  have all of the pages listed in the menu below with the names specified.  PLEASE keep all of your pages within your teams namespace. 
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</html>
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Instructor: ZHE LYU
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{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
 
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!align="center"|[[Team:UGA-Georgia|Home]]
 
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!align="center"|[[Team:UGA-Georgia/Team|Team]]
 
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!align="center"|[https://igem.org/Team.cgi?year=2013&team_name=UGA-Georgia Official Team Profile]
 
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!align="center"|[[Team:UGA-Georgia/Project|Project]]
 
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!align="center"|[[Team:UGA-Georgia/Parts|Parts Submitted to the Registry]]
 
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!align="center"|[[Team:UGA-Georgia/Modeling|Modeling]]
 
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!align="center"|[[Team:UGA-Georgia/Notebook|Notebook]]
 
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!align="center"|[[Team:UGA-Georgia/Safety|Safety]]
 
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!align="center"|[[Team:UGA-Georgia/Attributions|Attributions]]
 
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|}
 
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'''February 2013'''
 +
-Training on anaerobic skills, i.e. use of anaerobic glassware, gassing chamber,
 +
anaerobic chamber, etc.
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You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.
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'''March 2013'''
 +
 
 +
-Purification of GS, AT and GS+AT via revival of frozen stocks and plating of sub-cultures.
 +
 
 +
 
 +
'''April 2013'''
 +
 
 +
-Further continuation of purification of GS, AT and GS+AT via picking colonies, creating sub-cultures and plating.
 +
 
 +
-Test of Puromycin strength.
 +
 
 +
-Creation of frozen stocks of purified GS, AT and GS+AT transformants.
 +
 
 +
 
 +
'''June 2013'''
 +
 
 +
-Extraction of Geraniol from extra-cellular and intra-cellular content of samples and preparation for GC/MS evaluation
 +
 
 +
-Sequencing of pAW50-mCherry vector
 +
 
 +
-Analysis of pAW50-mCherry sequence
 +
 
 +
-Transformation of pAW50-mCherry vector into Methanococcus
 +
 
 +
-Testing Fluorescence of pAW50-mCherry
 +
 
 +
-Purification of pAW50-mCherry cultures via plating of transformants, and creating sub-cultures of colonies picked
 +
 
 +
-Creation of frozen stocks of pAW50-mCherry in Methanococcus
 +
 
 +
 
 +
'''July 2013'''
 +
 
 +
-Creation and analysis of the "killing & inhibiting" experiment where we test the maximum amount of product a 5ml culture of Methanococcus can tolerate in cultures with high OD (killing of grown cells) and low OD (inhibiting of growth).
 +
 
 +
-Innovation of an adapter to allow the use of syringe needles on micropipettes.
 +
 
 +
-Expanded upon the original extraction protocol for higher efficiency of extraction of geraniol from Methanococcus cultures.
 +
 
 +
- GC/MS Evaluation and analysis (see results tab)
 +
 
 +
 
 +
 
 +
'''August 2013'''
 +
 
 +
-Revival and PCR of all GS and AT frozen stocks to confirm insert.
 +
 
 +
--Extraction of Geraniol from extra-cellular and intra-cellular content of all GS frozen stocks and preparation for GC/MS evaluation
 +
 
 +
 
 +
'''September 2013'''
 +
 
 +
-Transformation of V4 into Methanococcus
 +
 
 +
-Testing Fluorescence of V4
 +
 
 +
-Purification of V4 cultures via plating of transformants, and creating sub-cultures of colonies picked
 +
 
 +
-Creation of frozen stocks of V4 in Methanococcus
 +
 
 +
-GC/MS Evaluation and analysis (see results tab)
 +
 
 +
== '''E. coli Lab''' ==
 +
 
 +
Instructor: Rachit Jain
 +
 
 +
 
 +
'''February 2013'''
 +
 
 +
-Training on PCR and heat shock transformation
 +
 
 +
 
 +
'''March 2013'''
 +
 
 +
-PCR of GS gene
 +
 
 +
-Cloning of GS gene into pAW-50.
 +
 
 +
-Transformation of pAW-50 GS into species XL1-Blue. Transformed species spread onto ampicillin plates
 +
 
 +
-Verification of GS transformants via digestion and gel verification
 +
 
 +
 
 +
'''April 2013'''
 +
 
 +
-Colonies for GS, AT, and GS+AT were selected from Methanococcus and corresponding genes were extracted. PCR and gel verification of GS, AT, and GS+AT genes. Results concluded successful in locating vectors from Methanococcus.
 +
 
 +
-PCR and gel verification of mCherry gene
 +
 
 +
-Cloning of mCherry gene into pAW-50 vector
 +
 
 +
-Verification of cloning via digestion and gel verification. After positive verification, ligation of digested products and heat shock transformation into XL1-Blue. XL1-Blue transformants spread onto ampicillin plates.
 +
 
 +
-XL1-Blue colonies picked and screened. Purification of pAW-50 mCherry from colonies, and stored in -20°C freezer.
 +
 
 +
 
 +
'''June 2013'''
 +
 
 +
-Transformation of pAW-50 mCherry into XL1-Blue and creation of stock
 +
 
 +
-PCR of two mCherry sequences: one without RBS sequence (V2) and one containing RBS sequence (V4). Gel Verification of tools 1 and 2.
 +
 
 +
-Digestion and cloning of V2 and V4 into pAW-50 mCherry vector
 +
 
 +
 
 +
'''July 2013'''
 +
 
 +
-Purification of digested vector, ligation, and transformation of vector into XL1-Blue. Transformants plated onto ampicillin plates.
 +
 
 +
-V2 and V4 colonies extracted and screened. Permanent stocks of V2B and V4B stored in -80°C freezer.

Latest revision as of 23:26, 22 September 2013

Notebook

Methanococcus Lab

Instructor: ZHE LYU


February 2013

-Training on anaerobic skills, i.e. use of anaerobic glassware, gassing chamber, anaerobic chamber, etc.


March 2013

-Purification of GS, AT and GS+AT via revival of frozen stocks and plating of sub-cultures.


April 2013

-Further continuation of purification of GS, AT and GS+AT via picking colonies, creating sub-cultures and plating.

-Test of Puromycin strength.

-Creation of frozen stocks of purified GS, AT and GS+AT transformants.


June 2013

-Extraction of Geraniol from extra-cellular and intra-cellular content of samples and preparation for GC/MS evaluation

-Sequencing of pAW50-mCherry vector

-Analysis of pAW50-mCherry sequence

-Transformation of pAW50-mCherry vector into Methanococcus

-Testing Fluorescence of pAW50-mCherry

-Purification of pAW50-mCherry cultures via plating of transformants, and creating sub-cultures of colonies picked

-Creation of frozen stocks of pAW50-mCherry in Methanococcus


July 2013

-Creation and analysis of the "killing & inhibiting" experiment where we test the maximum amount of product a 5ml culture of Methanococcus can tolerate in cultures with high OD (killing of grown cells) and low OD (inhibiting of growth).

-Innovation of an adapter to allow the use of syringe needles on micropipettes.

-Expanded upon the original extraction protocol for higher efficiency of extraction of geraniol from Methanococcus cultures.

- GC/MS Evaluation and analysis (see results tab)


August 2013

-Revival and PCR of all GS and AT frozen stocks to confirm insert.

--Extraction of Geraniol from extra-cellular and intra-cellular content of all GS frozen stocks and preparation for GC/MS evaluation


September 2013

-Transformation of V4 into Methanococcus

-Testing Fluorescence of V4

-Purification of V4 cultures via plating of transformants, and creating sub-cultures of colonies picked

-Creation of frozen stocks of V4 in Methanococcus

-GC/MS Evaluation and analysis (see results tab)

E. coli Lab

Instructor: Rachit Jain


February 2013

-Training on PCR and heat shock transformation


March 2013

-PCR of GS gene

-Cloning of GS gene into pAW-50.

-Transformation of pAW-50 GS into species XL1-Blue. Transformed species spread onto ampicillin plates

-Verification of GS transformants via digestion and gel verification


April 2013

-Colonies for GS, AT, and GS+AT were selected from Methanococcus and corresponding genes were extracted. PCR and gel verification of GS, AT, and GS+AT genes. Results concluded successful in locating vectors from Methanococcus.

-PCR and gel verification of mCherry gene

-Cloning of mCherry gene into pAW-50 vector

-Verification of cloning via digestion and gel verification. After positive verification, ligation of digested products and heat shock transformation into XL1-Blue. XL1-Blue transformants spread onto ampicillin plates.

-XL1-Blue colonies picked and screened. Purification of pAW-50 mCherry from colonies, and stored in -20°C freezer.


June 2013

-Transformation of pAW-50 mCherry into XL1-Blue and creation of stock

-PCR of two mCherry sequences: one without RBS sequence (V2) and one containing RBS sequence (V4). Gel Verification of tools 1 and 2.

-Digestion and cloning of V2 and V4 into pAW-50 mCherry vector


July 2013

-Purification of digested vector, ligation, and transformation of vector into XL1-Blue. Transformants plated onto ampicillin plates.

-V2 and V4 colonies extracted and screened. Permanent stocks of V2B and V4B stored in -80°C freezer.

Retrieved from "http://2013.igem.org/Team:UGA-Georgia/Notebook"