Team:Munich/chibiobrick2.html
7/31
Chi6 Oligodimerization
| Participants: | Domi |
| Protocol: | Oligodimerization |
8/1
assembling pSB1C3_Chi6
| Participants: | Domi |
| Protocol: | Restriction digest, PCR purify, Gibson assembly, chem.trafo |
| Notes: | XbaI, SpeI-HF; BB: 46 ng/µl |
| Results: |
8/2
PCR to assemble Chi6 oligomers
| Participants: | Enikö |
| Protocol: | PCR |
| Notes: | |
| Results: |
8/3
Gibson assembly to make pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | Gibson assembly |
| Notes: | |
| Results: |
8/3
Ligation to make pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | Ligation |
| Notes: | |
| Results: |
8/3
transforming DH5a with pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | chem trafo |
| Notes: | |
| Results: | no colonies |
8/6
DNA Mini prep from E. Coli Dh5a pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | mini prep |
| Notes: | |
| Results: | concentration |
8/6
Test Digest of pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | Restrictiondigest |
| Notes: | |
| Results: | no bands |
8/7
PCR to create Chi6 dimer
| Participants: | Enikö |
| Protocol: | PCR |
| Notes: | |
| Results: | no bands |
8/7
Digest Chi6 with EcoRI, SpeI
| Participants: | Enikö |
| Protocol: | Restrictiondigest |
| Notes: | |
| Results: | no bands |
8/7
PCR purify Chi6 digested with EcoRI, SpeI
| Participants: | Enikö |
| Protocol: | PCR purify |
| Notes: | |
| Results: |
8/7
Ligate pSB1C3 and Chi6 digested with EcoRI, SpeI
| Participants: | Enikö |
| Protocol: | Ligation |
| Notes: | |
| Results: | colonies |
8/8
DNA miniprep pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | miniprep |
| Notes: | |
| Results: | concentration |
8/8
sequencing pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | sequencing |
| Notes: | |
| Results: | ? |
8/9
sequencing pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | sequencing |
| Notes: | |
| Results: | ? |
8/12
PCR to assemble pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | PCR |
| Notes: | expected: 2kb |
| Results: | ? |
8/16
PCR to assemble pSB1C3_Chi6
| Participants: | Enikö |
| Protocol: | PCR |
| Notes: | because oligodimerisation & ligation failed, we decided to amplify chi6 by pcr; primers: Chi6_2clone & Chi6_fill cycle: 98° 2 min; 98° 20 s; 57° 30s; 72° 20s (to 2 5 x) 72° 2 min expect: ca 100 bp Backbone: BBS_AgeI_chi6_fw & BBP_SalI_Chi8_rv; AT: 62° expect: 2 kb |
| Results: | ? |
8/20
digest Chi6 to assemble pSB1C3_chi6
| Participants: | Enikö |
| Protocol: | restrictiondigest |
| Notes: | ageI, SalI |
| Results: | didnt work, ordered 3 different primer pairs |
8/30
preparing pSB1C3 backbone
| Participants: | Enikö |
| Protocol: | Restriction digest, Agarose gel, gelextraction |
| Notes: | EcoRi & SpeI |
| Results: | PIC |
8/30
dimerization of chi primers to assemble pSB1C3_chi6
| Participants: | Enikö |
| Protocol: | olidimerization, Agarose Gel, Gel extraction, Ligation, chem trafo |
| Notes: | |
| Results: | colonies |
9/2
Test-PCR of pSB1C3_chi6
| Participants: | Enikö |
| Protocol: | PCR |
| Notes: | VF2, VR; expected: 414bp |
| Results: | colonies |
9/5
Mini-prep of pSB1C3_chi6
| Participants: | Katja |
| Protocol: | mini prep |
| Notes: | |
| Results: |