Difference between revisions of "Part:BBa K2235009"

Revision as of 15:25, 26 October 2017

Sialidase composite with T7 promoter and RBS

Sialidase enzyme coding sequence with T7 promoter and RBS

Molecular cloning

Ligation of sialidase composite insert into pSB1C3

The gblock containing T7 promoter-RBS-sialidase was ligated into the Chloramphenicol plasmid backbone (pSB1C3). Preliminary confirmation of cloning was done by double digest Ecor1 and Pst1. Figure 1 represents the double digestion result: bands were visible at ~1600 bp and ~2000 bp which represents the insert and plasmid backbone respectively.

Figure 1: From left to right: M DNA ladder, next 2 lanes are plasmids digested with Ecor1 and Pst1.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 127
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 574
Illegal NgoMIV site found at 649
Illegal NgoMIV site found at 739
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1119

@@ Line 10: / Line 10: @@
 The gblock containing T7 promoter-RBS-sialidase was ligated into the Chloramphenicol plasmid backbone (pSB1C3). Preliminary confirmation of cloning was done by double digest Ecor1 and Pst1. Figure 1 represents the double digestion result: bands were visible at ~1600 bp and ~2000 bp which represents the insert and plasmid backbone respectively.
-[[File:SiaComp_1.png|600px|thumb|left|Figure 1]]
+[[File:SiaComp_1.png|600px|thumb|left|Figure 1:
+From left to right: M DNA ladder, next 2 lanes are plasmids digested with Ecor1 and Pst1.]]
-From left to right: M DNA ladder, next 2 lanes are plasmids digested with Ecor1 and Pst1.
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