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Team:Kyoto/CiC/Experiment
From 2009.igem.org
(→Construction) |
(→Construction) |
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†RBS+HIV-TAT+HIStag+(LALAAAA)3 was made by elongation of primer dimer. | †RBS+HIV-TAT+HIStag+(LALAAAA)3 was made by elongation of primer dimer. | ||
(Forward primer; | (Forward primer; | ||
| + | |||
cggaattcgcggccgcttctagagaaagaggagaaatactagATGTATGGACGTAAAAAACGTCGTGGACGTCGTCGTGGCGGCGGTCATCATCATCATCACCATGGCGG | cggaattcgcggccgcttctagagaaagaggagaaatactagATGTATGGACGTAAAAAACGTCGTGGACGTCGTCGTGGCGGCGGTCATCATCATCATCACCATGGCGG | ||
Reverse primer; | Reverse primer; | ||
| + | |||
ctgcagcggccgctactagtaTTACGCGGCCGCCGCCAGGGCCAGCGCGGCCGCCGCCAGGGCCAGCGCGGCCGCCGCCAGGGCCAGGCCACCGCCATGGTGATGATGATG) | ctgcagcggccgctactagtaTTACGCGGCCGCCGCCAGGGCCAGCGCGGCCGCCGCCAGGGCCAGCGCGGCCGCCGCCAGGGCCAGGCCACCGCCATGGTGATGATGATG) | ||
| Line 43: | Line 45: | ||
First-stage PCR | First-stage PCR | ||
Forward primer; | Forward primer; | ||
| + | |||
TTTAAACCGGCGACCCGTACCCTGTGCTCTTCTCGTTATCTGCTGcgtaaaggagaagaacttttcactggagttg | TTTAAACCGGCGACCCGTACCCTGTGCTCTTCTCGTTATCTGCTGcgtaaaggagaagaacttttcactggagttg | ||
| + | |||
Reverse primer; | Reverse primer; | ||
| + | |||
agtgagctgataccgctcgc | agtgagctgataccgctcgc | ||
Second-stage PCR | Second-stage PCR | ||
Forward primer; | Forward primer; | ||
| + | |||
cggaattcgcggccgcttctagagaaagaggagaaatactagATGCTGAGCCTGCGTCAGTCTATTCGTTTTTTTAAACCGGCGACCCGTAC | cggaattcgcggccgcttctagagaaagaggagaaatactagATGCTGAGCCTGCGTCAGTCTATTCGTTTTTTTAAACCGGCGACCCGTAC | ||
| + | |||
Reverse primer; | Reverse primer; | ||
| + | |||
agtgagctgataccgctcgc | agtgagctgataccgctcgc | ||
</div><!-- /#rightcontents --> | </div><!-- /#rightcontents --> | ||
Revision as of 05:23, 20 October 2009
Experiment
Construction
HIV-TAT::(LALAAAA)3 expressing vector
†RBS+HIV-TAT+HIStag+(LALAAAA)3 was made by elongation of primer dimer. (Forward primer;
cggaattcgcggccgcttctagagaaagaggagaaatactagATGTATGGACGTAAAAAACGTCGTGGACGTCGTCGTGGCGGCGGTCATCATCATCATCACCATGGCGG Reverse primer;
ctgcagcggccgctactagtaTTACGCGGCCGCCGCCAGGGCCAGCGCGGCCGCCGCCAGGGCCAGCGCGGCCGCCGCCAGGGCCAGGCCACCGCCATGGTGATGATGATG)
Signal for TIM23 complex::GFP expressing vector
†RBS+Signal for TIM23 complex::GFP+terminator was made by two-stage PCR.
First-stage PCR Forward primer;
TTTAAACCGGCGACCCGTACCCTGTGCTCTTCTCGTTATCTGCTGcgtaaaggagaagaacttttcactggagttg
Reverse primer;
agtgagctgataccgctcgc
Second-stage PCR Forward primer;
cggaattcgcggccgcttctagagaaagaggagaaatactagATGCTGAGCCTGCGTCAGTCTATTCGTTTTTTTAAACCGGCGACCCGTAC
Reverse primer;
agtgagctgataccgctcgc
