Team:SDU-Denmark/labnotes

From 2010.igem.org

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(Miniprep of BBa_K098995 in pSB1A2)
(Restriction digest of BBa_E0040 miniprep product)
 
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Nr. 28 in the freezer = K098995<br><br>
Nr. 28 in the freezer = K098995<br><br>
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=== Extraction of [http://partsregistry.org/Part:BBa_B0034 BBa_B0034] from pSB1A2 plasmid using PFU ===
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=== Extraction of [http://partsregistry.org/Part:BBa_B0034 B0034] from pSB1A2 plasmid using PFU ===
==== Preliminary PCR to amplify VF2-VR piece containing part and restriction sites ====
==== Preliminary PCR to amplify VF2-VR piece containing part and restriction sites ====
Date: July 14th <br>
Date: July 14th <br>
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--[[User:CKurtzhals|CKurtzhals]] 16:07, 15 July 2010 (UTC)
--[[User:CKurtzhals|CKurtzhals]] 16:07, 15 July 2010 (UTC)
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=== Miniprep and transformation of [http://partsregistry.org/Part:BBa_K274210 BBa_K274210] in pSB1A2 ===
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=== Miniprep and transformation of [http://partsregistry.org/Part:BBa_K274210 K274210] in pSB1A2 ===
Start date: July 14.th <br>
Start date: July 14.th <br>
''Methods:''  Fermentas GeneJET plasmid miniprep kit <br><br>
''Methods:''  Fermentas GeneJET plasmid miniprep kit <br><br>
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'''Transformation of the miniprepped plasmid:'''<br><br>
'''Transformation of the miniprepped plasmid:'''<br><br>
''Experiment done by:'' LC, Maria <br><br>  ''Protocol:'' TR 1.1 <br><br>
''Experiment done by:'' LC, Maria <br><br>  ''Protocol:'' TR 1.1 <br><br>
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''Notes:'' Transformed BBa_K274210 in pSB1A2 on LA plates with 100 microgram/ml ampicillin into ''E. coli'' MG1655. We used 2 microliter from the miniprep product for each sample.<br>
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''Notes:'' Transformed K274210 in pSB1A2 on LA plates with 100 microgram/ml ampicillin into ''E. coli'' MG1655. We used 2 microliter from the miniprep product for each sample.<br>
Our suspicion that the agar plates we made are faulty got confirmed, three more plates were destroyed while spreading the bacteria on them (Sample 1a, 2b, 2pellet and the positive control)<br><br>
Our suspicion that the agar plates we made are faulty got confirmed, three more plates were destroyed while spreading the bacteria on them (Sample 1a, 2b, 2pellet and the positive control)<br><br>
''Results:'' Cells grew just fine and the negative control was as expected. After a night at 37° the cells seemed a little more yellowish to the eye, than the usual. <br><br>
''Results:'' Cells grew just fine and the negative control was as expected. After a night at 37° the cells seemed a little more yellowish to the eye, than the usual. <br><br>
''Analysis:'' The transformations succeeded. <br><br>
''Analysis:'' The transformations succeeded. <br><br>
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=== Coloni PCR of pSB1A2 w. [http://partsregistry.org/Part:BBa_E0040 BBa_E0040] (transformation 13/7) ===
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=== Coloni PCR of pSB1A2 w. [http://partsregistry.org/Part:BBa_E0040 E0040] (transformation 13/7) ===
Start date: July 14th <br>
Start date: July 14th <br>
''Methods:''  Taq coloni PCR and gel electrophoresis <br><br>
''Methods:''  Taq coloni PCR and gel electrophoresis <br><br>
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--[[User:Tipi|Tipi]] 16:09, 19 July 2010 (UTC)
--[[User:Tipi|Tipi]] 16:09, 19 July 2010 (UTC)
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=== Transformation of [http://partsregistry.org/Part:BBa_K081005 BBa_K081005] in pSB1A2 (constitutive promoter and RBS combined) and [http://partsregistry.org/Part:BBa_R0011 BBa_R0011] in pSB1A2 in Top 10 E.Coli ===
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=== Transformation of [http://partsregistry.org/Part:BBa_K081005 K081005] in pSB1A2 (constitutive promoter and RBS combined) and [http://partsregistry.org/Part:BBa_R0011 R0011] in pSB1A2 in Top 10 E.Coli ===
Start date: July 15th - 16th <br>
Start date: July 15th - 16th <br>
''Methods:''  Overnight culture, making competent cells, transformation <br><br>
''Methods:''  Overnight culture, making competent cells, transformation <br><br>
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--[[User:Lclund|Lclund]] 10:00, 19 July 2010 (UTC)
--[[User:Lclund|Lclund]] 10:00, 19 July 2010 (UTC)
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==== Restriction digest of [http://partsregistry.org/Part:BBa_E0040 BBa_E0040] miniprep product ====
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==== Restriction digest of [http://partsregistry.org/Part:BBa_E0040 E0040] miniprep product ====
Start date: July 16th<br>
Start date: July 16th<br>
''Methods:''  Restriction digest <br><br>
''Methods:''  Restriction digest <br><br>
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<br><br>
<br><br>
''Analysis:'' Sample 2 and 3 got pooled and frozen, 1 and 4 were discarded, since their length didn't seem exactly right.<br><br>
''Analysis:'' Sample 2 and 3 got pooled and frozen, 1 and 4 were discarded, since their length didn't seem exactly right.<br><br>
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Nr. 12 in the freezer = BBa_E0040 miniprep product in pSB1A2.<br>
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Nr. 12 in the freezer = E0040 miniprep product in pSB1A2.<br>
--[[User:Lclund|Lclund]] 10:00, 19 July 2010 (UTC)
--[[User:Lclund|Lclund]] 10:00, 19 July 2010 (UTC)

Latest revision as of 19:16, 24 October 2010