Difference between revisions of "Part:BBa K1789015"
Line 19: Line 19:
  
 
Functional check:
 
Functional check:
−
  Methodology: In order to investigate whether the fluorescence
+
Methodology: In order to investigate whether the fluorescence
−
observed was due to the presence of the original GFP-N2 construction. First of all, the recombinant colonies were transferred into LB culture medium with chloramphenicol, and shook overnight at 37 ℃, then took the medium for PCR templet. Meanwhile, we extracted plasmid DNA using a miniprep kit and did a digest with Xbal1 and Pst1 to check for the presence of the original insert and size of the unfolded vector, respectively.
+
observed was due to the presence of the original GFP-N2 construction.  
−
  Result and discussion: The DNA positive strand and the insert is very clearly visible at just below 3kb and 5kb. This proves the presence of GFP-N2 in culture.
+
First of all, the recombinant colonies were transferred into  
 +
LB culture medium with chloramphenicol, and shook overnight at 37 ℃,  
 +
then took the medium for PCR templet. Meanwhile, we extracted plasmid DNA  
 +
using a miniprep kit and did a digest with Xbal1 and Pst1 to check  
 +
for the presence of the original insert and size of the unfolded vector, respectively.
 +
Result and discussion: The DNA positive strand and the insert  
 +
is very clearly visible at just below 3kb and 5kb.  
 +
This proves the presence of GFP-N2 in culture.

Revision as of 09:32, 18 September 2015

GFP_N1

This device is a negative control of GFP with scaf.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2311
    Illegal BamHI site found at 4908
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1304
    Illegal BsaI.rc site found at 3390
    Illegal BsaI.rc site found at 3798
    Illegal BsaI.rc site found at 4104
    Illegal BsaI.rc site found at 5094



Functional check: Methodology: In order to investigate whether the fluorescence observed was due to the presence of the original GFP-N2 construction. First of all, the recombinant colonies were transferred into LB culture medium with chloramphenicol, and shook overnight at 37 ℃, then took the medium for PCR templet. Meanwhile, we extracted plasmid DNA using a miniprep kit and did a digest with Xbal1 and Pst1 to check for the presence of the original insert and size of the unfolded vector, respectively. Result and discussion: The DNA positive strand and the insert is very clearly visible at just below 3kb and 5kb. This proves the presence of GFP-N2 in culture.