Team:HokkaidoU Japan/Notebook/Promoter WeakPromoter-I732020

From 2013.igem.org

Maestro E.coli

Notebook

Weak Promoter-I732020

Start
Transformation
- Fri Jun 7
J23112, J23103, J23117, J23106, J23111 from distribution kit
each of 1µL DNA to DH5α
Culture
- Sat Jun 8 and Fri Jun 21
J23112, J23103, J23117, J23106, J23111
I732020 from our stock
Mini-prep
- Sun Jun 9 and Sat Jun 22
J23112, J23103, J23117, J23106, J23111, I732020
PCR
- Wed Jun 26
I732020
100up, 200dn, KOD Plus Neo
Digestion
- Mon Jun 10, Sat Jun 22 and Wed Jun 26
Cut (J23112, J23103, J23117, J23106, J23111) with SpeI, PstI (using 10×H buffer)
Cut I732020 with XbaI, PstI (using 10×M buffer)
Gel extraction
- Tue Jun 11 and Mon Jun 24
J23112, J23103, J23117, J23106, J23111
Antarctic phosphatase
- Sun Jun 30
J23112, J23103, J23117, J23106, J23111
Ligation
- Tue Jun 11, Mon Jun 24 and Sun Jun 30
Ligate (J23112, J23103, J23117, J23106, J23111) with I732020
Transformation
- Tue Jun 11, Mon Jun 24 and Sun Jun 30
(J23112, J23103, J23117, J23106, J23111)-I732020
each of 1µL DNA to DH5α
Colony PCR & Culture
- Sun Jun 30
(J23112, J23103, J23117, J23106, J23111)-I732020
Mini-prep
- Mon Jul 1
(J23112, J23103, J23117, J23106, J23111)-I732020
PCR
- Fri Jul 5
(J23112, J23103, J23117, J23106, J23111)-I732020
EX-f, PS-r, KOD FX NEO
PCR purification
- Sat Jul 6
(J23112, J23103, J23117, J23106, J23111)-I732020
Digestion
- Sat Jul 6
Cut K1084007 on pSB1C3 with EcoRI, PstI (using 10×H buffer)
Cut (J23112, J23103, J23117, J23106, J23111)-I732020 with EcoRI, PstI (using 10×H buffer)
Antarctic phophatase
- Sun Jul 7
pSB1C3
Ligation
- Sun Jul 7
Ligate pSB1C3 with (J23112, J23103, J23117, J23106, J23111)-I732020
Transformation
- Sun Jul 7
(J23112, J23103, J23117, J23106, J23111)-I732020
Complete!

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