Team:BIOTEC Dresden/Notebook2-1
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+ | '''1st September‘09:''' | ||
+ | |||
+ | 1. Inoculation of 5 colonies from pTetFlp-KanR plasmid containing bacteria is sub-cultured into liquid agar and solid medium for overnight growth. | ||
+ | |||
+ | 2. Minipreps of pRhaFlp-CmR clones has been done. | ||
+ | |||
+ | 3. A Red/ET recombination has been done to insert PCR-CmR into pSC101-pRhaFlp plasmid and plated on Cm plates for overnight incubation. | ||
+ | |||
+ | '''2nd September‘09:''' | ||
+ | |||
+ | 1. The concentrations of pRhaFlp-CmR miniprep DNAs has been done using nanodrop measuring device. | ||
+ | |||
+ | 2. A restriction digest of parental pRhaFlp and pTetFlp plasmids along with pRhaFlp-CmR has been done with EcoRI and other minipreps of pRhaFlp-CmR with PstI. | ||
+ | |||
+ | 3. Agarose Gel Electrophoresis is done to check the digests. | ||
+ | |||
+ | '''3rd September‘09:''' | ||
+ | |||
+ | 1. A sequencing reaction has been set up for the PCR product of the plasmid pR6K-AmpSpec with Spec-forward and Spec-reverse primers using Taq polymerase. | ||
+ | |||
+ | 2. Colonies from the pTetFlp-KanR are sub-cultured on new trimethoprin plates. | ||
+ | |||
+ | 3. From Red/ET recombination of pRhaFlp clones positive colonies are reincubated from induced plate. | ||
+ | |||
+ | '''4th September‘09:''' | ||
+ | |||
+ | 1. Minipreps of pRhaFlp-CmR clones has been done from the overnight cultures. | ||
+ | |||
+ | 2. Re-electroporation of minipreps of 1, 12 and 13 of pRhaFlp-CmR has been done. | ||
+ | |||
+ | |||
{{:Team:BIOTEC_Dresden/NewTemplateEnd}} | {{:Team:BIOTEC_Dresden/NewTemplateEnd}} |
Latest revision as of 12:16, 21 October 2009
1st September‘09:
1. Inoculation of 5 colonies from pTetFlp-KanR plasmid containing bacteria is sub-cultured into liquid agar and solid medium for overnight growth.
2. Minipreps of pRhaFlp-CmR clones has been done.
3. A Red/ET recombination has been done to insert PCR-CmR into pSC101-pRhaFlp plasmid and plated on Cm plates for overnight incubation.
2nd September‘09:
1. The concentrations of pRhaFlp-CmR miniprep DNAs has been done using nanodrop measuring device.
2. A restriction digest of parental pRhaFlp and pTetFlp plasmids along with pRhaFlp-CmR has been done with EcoRI and other minipreps of pRhaFlp-CmR with PstI.
3. Agarose Gel Electrophoresis is done to check the digests.
3rd September‘09:
1. A sequencing reaction has been set up for the PCR product of the plasmid pR6K-AmpSpec with Spec-forward and Spec-reverse primers using Taq polymerase.
2. Colonies from the pTetFlp-KanR are sub-cultured on new trimethoprin plates.
3. From Red/ET recombination of pRhaFlp clones positive colonies are reincubated from induced plate.
4th September‘09:
1. Minipreps of pRhaFlp-CmR clones has been done from the overnight cultures.
2. Re-electroporation of minipreps of 1, 12 and 13 of pRhaFlp-CmR has been done.