Difference between revisions of "Part:BBa K2762007"

Line 7: Line 7:
  
 
==Characterization==
 
==Characterization==
===Colony PCR of finished construct===
+
===SDS-PAGE of PRK===
 +
To find out whether the gene prk is successfully expressed in E. coli, we conducted a SDS-PAGE test. The cells were harvested by centrifuging at 10,000×g for 10 min., and then washed with deionized water for 2 times. The cell density was adjusted to an OD600 of 5 as the sample of whole cell (WC, whole cell catalyst).
 +
Finally, WC was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with 10% separating gel and 4% stacking gel. Proteins were visualized by staining with Coomassie blue R-250 and were scanned with an Image scanner.
 +
The result is shown below.
 +
 
  
  

Revision as of 06:31, 29 September 2018


PlacI-B0034-prk-B0015

Usage and Biology

Characterization

SDS-PAGE of PRK

To find out whether the gene prk is successfully expressed in E. coli, we conducted a SDS-PAGE test. The cells were harvested by centrifuging at 10,000×g for 10 min., and then washed with deionized water for 2 times. The cell density was adjusted to an OD600 of 5 as the sample of whole cell (WC, whole cell catalyst).

Finally, WC was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with 10% separating gel and 4% stacking gel. Proteins were visualized by staining with Coomassie blue R-250 and were scanned with an Image scanner.

The result is shown below.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]