Difference between revisions of "Part:BBa K1926013:Design"
 
Line 8: Line 8:
 
The mCherry with tags and RTSs are constructed using PCR and oligonucleotide ligation by the following procedure. Our UNITs all contain XbaI and SpeI enzyme sites, so we can easily line up the UNITs using 3A assembly.   
 
The mCherry with tags and RTSs are constructed using PCR and oligonucleotide ligation by the following procedure. Our UNITs all contain XbaI and SpeI enzyme sites, so we can easily line up the UNITs using 3A assembly.   
  
−
[[File:5-连接过程改.png|800px|thumb|left|'''Figure 1:''' Three steps to construct our UNITs using PCR and oligonucleotide ligation.]]
+
[[File:5-连接过程改.png|600px|thumb|left|'''Figure 1:''' Three steps to construct our UNITs using PCR and oligonucleotide ligation.]]
 
<br style="clear: both" />
 
<br style="clear: both" />
  

Latest revision as of 07:31, 19 October 2016

The mCHERRY UNIT: mCherry flanked by Vox


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]


Design Notes

The mCherry with tags and RTSs are constructed using PCR and oligonucleotide ligation by the following procedure. Our UNITs all contain XbaI and SpeI enzyme sites, so we can easily line up the UNITs using 3A assembly.

Figure 1: Three steps to construct our UNITs using PCR and oligonucleotide ligation.



Source

The sequence of Vox and DBOX was retrieved from Addgene and got through denovo synthesis. mCHERRY and sv40 terminator were got from commercial plasmid: pentry dcas9 mcherry bira and pentry nls GFP/pcdna3.1, respectfully.


References

1. Voutev, R. and E.J. Hubbard, A "FLP-Out" system for controlled gene expression in Caenorhabditis elegans. Genetics, 2008. 180(1): p. 103-19.