Difference between revisions of "Part:BBa K3790151"
TiTong Fudan (Talk | contribs) (→Experimental Results) |
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[[File:T--Fudan--ccic8-transparent-logo.png|100px|right|2021 Fudan]] | [[File:T--Fudan--ccic8-transparent-logo.png|100px|right|2021 Fudan]] | ||
− | The fusion protein obtained by linking the C-terminus of Bst to the N- | + | The fusion protein obtained by linking the C-terminus of Bst to the N-terminus of S.ssb via a (G2S)3 linker. |
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===Experimental Results=== | ===Experimental Results=== | ||
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The length of Bst-(G2S)3-S.ssb DNA was 2235 bp, which is approximately 2250 bp after adding homology arms to both ends for PCR cloning. We isolated the target DNA by gel extraction for subsequent reactions. | The length of Bst-(G2S)3-S.ssb DNA was 2235 bp, which is approximately 2250 bp after adding homology arms to both ends for PCR cloning. We isolated the target DNA by gel extraction for subsequent reactions. | ||
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Mini-prep was performed the following day, and the resulting plasmids were sent to Sanger sequencing. | Mini-prep was performed the following day, and the resulting plasmids were sent to Sanger sequencing. | ||
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− | <span class='h3bb'>Sequence and Features</span> | + | <span class='h3bb'>'''Sequence and Features'''</span> |
<partinfo>BBa_K3790151 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3790151 SequenceAndFeatures</partinfo> | ||
Latest revision as of 20:38, 21 October 2021
Bst-(G2S)3-S.ssb
Introduction
The fusion protein obtained by linking the C-terminus of Bst to the N-terminus of S.ssb via a (G2S)3 linker.
Contents
Experimental Results
The length of Bst-(G2S)3-S.ssb DNA was 2235 bp, which is approximately 2250 bp after adding homology arms to both ends for PCR cloning. We isolated the target DNA by gel extraction for subsequent reactions.
After ClonExpress homologous recombination reaction, Fast-T1 competent bacteria were transformed with reactants, and spread onto LB plates with antibiotics. We picked colony after 16 h culturing plates at 37℃. We further grow clones in 2 ml LB liquid medium with antibiotics, 37℃ shaking overnight.
Mini-prep was performed the following day, and the resulting plasmids were sent to Sanger sequencing.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]