Team:Heidelberg/Notebook/Sensing Group/Notebook/3rdweek

(Difference between revisions)

Revision as of 22:54, 26 October 2008

PCR again for LuxQ-1 and LuxQ-2 for Fusion constructs. Different Mg concentrations from 1% to 4% were used. For LuxQ-2 another PCR was run using as template V. harveyi colonies

PCR of LuxQ-1 and LuxQ-2 for subsequent fusion PCR

Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar mixed with 1 µl LuxQ)
preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep
digestion of pDK48 with NcoI/NdeI
digestion of pTrc99alpha and LuxQ (with BamHI and NcoI) and gel extraction

Miniprep of pDK48 and LuxS
gel of Fusion constructs and gelextraction of pDK48 from yesterday
afterwards another gel was run to purify the Fusion constructs via gelextraction

Fusion constructs and pDK48 gel extraction

@@ Line 124: / Line 124: @@
 == Monday, 08/18/2008 ==
-* PCR for LuxQ-1 and LuxQ-2 for Fusion constructs. Different Mg concentrations between 1-4 % were used. For LuxQ-2 another PCR was run using as template ''V. harveyi'' colonies
+* PCR again for LuxQ-1 and LuxQ-2 for Fusion constructs. Different Mg concentrations from 1% to 4% were used. For LuxQ-2 another PCR was run using as template ''V. harveyi'' colonies
 [[Image:HD 080818-F1 LuxQ1 2.png|center|thumb|500px|PCR of LuxQ-1 and LuxQ-2 for subsequent fusion PCR]]
-* Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar, 1 µl LuxQ)
+* Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar mixed with 1 µl LuxQ)
 * preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep
 * digestion of pDK48 with NcoI/NdeI
-* digestion of pTrc99alpha and LuxQ (with BamHI and NcoI) and gelextraction
+* digestion of pTrc99alpha and LuxQ (with BamHI and NcoI) and gel extraction
 == Tuesday, 08/19/2008 ==