Difference between revisions of "Part:BBa K1431101:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | Design the primers and PCR by Q5<sup>®</sup> High-Fidelity DNA Polymerases from plasmid.<br> | + | Design the primers and PCR by Q5<sup>®</sup> High-Fidelity DNA Polymerases from plasmid. |
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| + | <center>https://static.igem.org/mediawiki/parts/9/9b/Plasmid_transfected.jpg</center> | ||
| + | <center>'''pBX-093 PB5-HS4-SV40-puro-2A-tetON3G-pA-HS4-TRE-AzaminGreen-2A-T'''</center><br> | ||
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Primer Forward: T TCTAGA TGGGATCAAGACTGGACAAGA<br> | Primer Forward: T TCTAGA TGGGATCAAGACTGGACAAGA<br> | ||
Primer Reverse: AAAACTGCAG CGGCCGC T ACTAGT A CCGAAGCCCAACCTTTCATA<br> | Primer Reverse: AAAACTGCAG CGGCCGC T ACTAGT A CCGAAGCCCAACCTTTCATA<br> | ||
Revision as of 03:25, 18 October 2014
TetOn-3G, an ideal controller of mammalian gene expression with TRE-3G promoter+PolyA
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Design the primers and PCR by Q5® High-Fidelity DNA Polymerases from plasmid.
Primer Forward: T TCTAGA TGGGATCAAGACTGGACAAGA
Primer Reverse: AAAACTGCAG CGGCCGC T ACTAGT A CCGAAGCCCAACCTTTCATA
Sequencing Results
We sent fresh bacteria broth for sequencing using standard Biobricks sequencing primer VF2/VR. The sequencing cooperation we used is Invitrogen Guangzhou filiale.
Sequence of sequencing primer we used:
VF2: tgccacctgacgtctaagaa
VR: attaccgcctttgagtgagc
The result shows the same sequence with our ideal design.
Source
The plasmid was from our instructor, Huangwei's lab. And we design primers to copy down by PCR.