Team:Tongji China/Record
Tongji iGEM
TongJi iGEM
Record
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Experiment
Plasmid Construction Parts (Week 1-6: June 5-July 10)
Week1
June 5
pUAST-3xHA transform into E.coli.
June 6
1. Pick up the single colony, and culture it with 10 mL 2xYT medium overnight.
2. Design sequencing primers of pUAST-3xHA.
June 7
1. Midi extraction of plasmids pUAST-3xHA.
2. Send it to GENEWIZ to sequence.
June 8
1. Verify the sequencing result, it is correct.
2. Digest pUAST-3xHA with BamHI overnight.
June 9
Extraction of Drosophila genome and RNA, purify the digested vector.
pUAST-3xHA transform into E.coli.
June 6
1. Pick up the single colony, and culture it with 10 mL 2xYT medium overnight.
2. Design sequencing primers of pUAST-3xHA.
June 7
1. Midi extraction of plasmids pUAST-3xHA.
2. Send it to GENEWIZ to sequence.
June 8
1. Verify the sequencing result, it is correct.
2. Digest pUAST-3xHA with BamHI overnight.
June 9
Extraction of Drosophila genome and RNA, purify the digested vector.
Week2
June 12
Reverse transcription of Drosophila RNA, then get the cDNA.
June 13
Design the primers for fragments’ PCR and fragments’ sequencing.
June 14
1. Clone pleP, Gal4, Gal80ts from genome (success) and TH from cDNA (failed), purify them.
2. Send them to GENEWIZ to sequence.
June 15
1. Verify the sequencing result, it is correct.
2. Ligase each of them with pLB vector and transform into E.coli.
3. Clone TH from cDNA again (failed).
June 16
1. Pick up the single colony, do bacteria liquid PCR.
2. Send 5 positive colony to GENEWIZ to sequence.
3. Design the In-Fusion primers.
4. Clone TH from cDNA again (failed).
Reverse transcription of Drosophila RNA, then get the cDNA.
June 13
Design the primers for fragments’ PCR and fragments’ sequencing.
June 14
1. Clone pleP, Gal4, Gal80ts from genome (success) and TH from cDNA (failed), purify them.
2. Send them to GENEWIZ to sequence.
June 15
1. Verify the sequencing result, it is correct.
2. Ligase each of them with pLB vector and transform into E.coli.
3. Clone TH from cDNA again (failed).
June 16
1. Pick up the single colony, do bacteria liquid PCR.
2. Send 5 positive colony to GENEWIZ to sequence.
3. Design the In-Fusion primers.
4. Clone TH from cDNA again (failed).
