| DATE |
LAB NOTE |
RESULT |
| 2015/3-4 |
Discussed project |
Wonderful |
| 2015/4/5 |
Made DH5α competent cell |
Succeed |
| 2015/4/8 |
TransformedBBa_R0082(Promoter (OmpR, positive)) |
Succeed |
| 2015/4/10 |
TransformedBBa_K592018 (cph8 with strong RB S),BBa_P0451(strongRBS:B0034+ cI (lambda): C0051+double terminator:B0010、B0012),BBa_C0040 (tetracycline repressor from transposon Tn10 (+LVA)) |
Succeed |
| 2015/4/15 |
TransformedBBa_K592016 (B0034-YF1-B0034-FixJ, blue light sensor and RR with RBS) , BBa_K823006(constitutive promoter) and BBa_K592006(FixK2 promoter) |
Succeed |
| 2015/4/2-4/17 |
Askedfor the cas9 part from other universities |
Failed |
| 2015/4/10-18 |
Askedfor the cas9 gene at our college and finally get from postgraduate |
Succeed |
| 2015/4/20 |
Made competent cell DH5α |
Succeed |
| 2015/4/25 |
TransformedBBa_S03518 (B0034-TetR); |
Succeed |
| 2015/4/28 |
TransformedBBa_S03877(Strong RBS+hol), BBa_K081021(B003+Pcya+ terminator) |
Succeed |
| 2015/4/29 |
TransformedBBa_B0015 (Double terminator，forward： B0010+B0012) |
Succeed |
| 2015/5/2 |
Got Hfr straIn; PCR cas9 sequence and attempted to ligated cas9 on pSB1C3 |
Succeed |
| 2015/5/3 |
Made DH5αcompetent cell |
Succeed |
| 2015/5/4 |
PCR tested the existence of Orit and TraI gene in Hfr; Double restriction digestedBBa_S03877and BBa_K081021；ligateddigestedBBa_S03877 and digestedBBa_K081021 |
Succeed |
| 2015/5/5 |
Transformed the product of ligation on the former day into DH5α( the successful product was marked as holpt) |
Succeed |
| 2015/5/7-8 |
T-A colony of OriT from Hfr |
Failed |
| 2015/5/10 |
Double restriction digestedholpt and BBa_R0082;ligateddigestedholpt and BBa_R0082 |
Succeed |
| 2015/5/11 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as holpto) |
Succeed |
| 2015/5/11-12 |
T-A colony of OriT from Hfr |
Succeed |
| 2015/5/14 |
Test the conjugation efficiency between Hfr and BL21 |
Succeed |
| 2015/5/19 |
Tired to ligated cas9 with constitutive promoter ;Double restriction digestedBBa_S03518 and BBa_B0015 ;ligateddigestedBBa_S03518 and BBa_B0015 |
Ligation failed; digestedion succeed |
| 2015/5/20 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as tetT) |
Succeed |
| 2015/5/24 |
T-A colony of TraI from Hfr ; Double restriction digestedBBa_R0082 and holpto;ligateddigestedBBa_R0082 and holpto |
T-A colonyfailed; restriction digestedionsucceed |
| 2015/5/25 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as cph8A) |
Succeed |
| 2015/5/28 |
Double restriction digestedtetT and BBa_C0040;ligateddigestedtetT and BBa_C0040 |
Succeed |
| 2015/5/29 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as tetra) |
Succeed |
| 2015/5/9-6/1 |
Overlap PCR to knock out TraI gene in Hfr |
Failed |
| 2015/6/2 |
T-A colony of TraI from Hfr; TransformedBBa_I13504 (reporter 1.0RBS+GFP+Terminator) |
T-A colony failed; Transform edation succeed |
| 2015/6/3 |
Made DH5α competent cell |
Succeed |
| 2015/6/4 |
Double restriction digestedcph8A , tetra and BBa_I13504;ligated digeste dcph8A and BBa_I13504, , tetra and BBa_I13504 |
Succeed |
| 2015/6/5 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as cpg and tetRg) |
Succeed |
| 2015/6/9 |
Double restriction digestedcph8A and tetRg;ligateddigestedcph8A and tetRg |
Succeed |
| 2015/6/10 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as ctg) |
Succeed |
| 2015/6/12 |
T-A colony of TraI from Hfr |
Failed |
| 2015/6/19 |
Double restriction digestedBBa_K592016 (B0034-YF1-B0034-FixJ, blue light sensor and RR with RBS) and BBa_B0015 (Double terminator，forward： B0010+B0012); ligateddigestedBBa_K592016 and BBa_B0015 |
Succeed |
| 2015/6/20 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as NT) |
Succeed |
| 2015/6/20 |
Made DH5αcompetent cell |
Succeed |
| 2015/6/15-6/21 |
DNA synthesis of promoter λ(the DNA is called Pci) |
Succeed |
| 2015/6/25 |
Double restriction digested NT,Pci , BBa_I13504and BBa_K592006;ligateddigested NT and BBa_K592006, Pci and BBa_K592006, Pci and BBa_I13504 |
Succeed |
| 2015/6/26 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as NG,GCi and Cig) |
Succeed |
| 2015/7/1 |
Double restriction digested NG and BBa_K823006;ligateddigested NG and BBa_K823006 |
Succeed |
| 2015/7/2 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as cngg) |
Succeed |
| 2015/7/4 |
Transformed the plasmid pET28a into DH5α |
Succeed |
| 2015/7/6 |
Double restriction digestedpET28a and cngg；ligateddigestedpET28a and cngg |
Succeed |
| 2015/7/7 |
Transformed the product of ligation on the former day into DH5α (the successful product was marked as pETcngg) |
Succeed |
| 2015/7/8 |
Made BL21 competent cell |
Succeed |
| 2015/7/10 |
Transformed the plasmid pETcngg into Made BL21 |
Succeed |
| 2015/7/13 |
Double restriction digestedcpg and Pet28a;ligateddigestedcpg and Pet28a |
Succeed |
| 2015/7/14 |
Transformed the product of ligation on the former day into DH5α |
Failed |
| 2015/7/15 |
Made DH5αcompetent cell |
Succeed |
| 2015/6/5-7/15 |
TraIgene knock out in Genescript company |
Succeed |
| 2015/7/17 |
Transformed the product of ligation on the former day into DH5α |
Failed |
| 2015/7/20 |
Double restriction digestedcpg, ctg and BBa_K823006 |
Succeed |
| 2015/7/16-7/20 |
Test the conjugation efficiency between Hfr and BL21 and between TraIknocked-out Hfr and DH5α |
Succeed |
| 2015/7/21 |
Transformed the product of ligation on the former day into DH5α |
Failed |
| 2015/7/23 |
Double restriction digestedcpg, ctg and BBa_K823006 |
Succeed |
| 2015/7/24 |
Transformed the product of ligation on the former day into DH5α |
Failed |
| 2015/7/25-8/4 |
Synthetized “GFPsg+terminater+constitutive promoter+1.0RBS” sequence by company. |
Succeed |
| 2015/8/5 |
Made DH5αcompetent cell ; Ligatedconstitutive promoter with GFP |
Succeed |
| 2015/8/7 |
Twice single restriction digestedcpg, ctg and Pet28a;ligateddigestedcpg and Pet28a; ctg and Pet28a; Ligated cas9 on pMD-18T |
Succeed |
| 2015/8/1-8/7 |
Verified constructions of three plasmids needed for InterLab Study |
Succeed |
| 2015/8/8 |
Transformed the product of ligation on the former day into DH5α |
Failed |
| 2015/8/10 |
Triedto ligate “GFPsg+terminater+ constitutive promoter+1.0RBS” sequence with cas9 |
Failed |
| 2015/8/10-8/12 |
InterLab Study read florescence intensity: First attempt |
Failed |
| 2015/8/13-8/15 |
InterLab Study read florescence intensity: second attempt |
Succeed |
| 2015/8/15 |
Double restriction digested cpg and Pet28a;ligated digested cpg and Pet28a |
Succeed |
| 2015/8/16 |
Transformed the product of ligation on the former day into DH5α |
Failed |
| 2015/8/20 |
Ligatedcas9on pet-28a. |
Failed |
| 2015/8/21 |
Twice single restriction digested cpg, ctg and Pet28a;ligated digested cpg and Pet28a; ctg and Pet28a; Ligated cas9 on pMD-18T |
Succeed |
| 2015/8/22 |
Transformed the product of ligation on the former day into DH5α |
Failed |
| 2015/8/21 |
T-A colony of TraI from Hfr |
Succeed |
| 2015/8/22 |
Verification of T-A colony |
Succeed |
| 2015/8/23 |
DNA sequencing of traI |
Failed |
| 2015/8/24-27 |
T-A colony of TraI from Hfr again |
Succeed |
| 2015/8/28 |
Twice single restriction digested traI and pSB1C3; ligated traI and pSB1C3. |
Failed |
| 2015/9/1-3 |
Twice single restriction digested traI and pSB1C3; ligated traI and pSB1C3. |
Succeed |
| 2015/9/4-7 |
Functional verification of traI |
Succeed |
| 2015/9/9 |
Another T-A colony of OriT from Hfr |
Succeed |
| 2015/9/10 |
Made DH5αcompetent cell |
Succeed |
| 2015/9/10 |
Submit the biobrick |
Succeed |
