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Harvard BioDesign/23 July 2014
From 2014.igem.org
(Created page with "Sequencing results were not good The plasmid did not have the insert") |
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Sequencing results were not good | Sequencing results were not good | ||
The plasmid did not have the insert | The plasmid did not have the insert | ||
| + | |||
| + | Transformed Gibson product (pBbE1a- CsgA- Spy Tag) into Mach I cells ( attempt 2) | ||
| + | Tried varying forms of Gibson product | ||
| + | 2 transformations add 2 microliters of Gibson product (1 & 3) | ||
| + | 2 transformations adding 4 microliters of Gibson product (2 & 4) | ||
| + | Used different transformation tubes → green tops | ||
| + | Transformation 1 was probably not good because I was not on the correct setting | ||
| + | Transformations 2 and 3 arc’ed first | ||
| + | Tried to send pulse again for 2 ---> a good read came up on the equipment | ||
| + | Tried to send pulse again for 3----> arc’ed once again | ||
| + | Transformation 4 went through without arcing | ||
| + | |||
| + | Grew up another colony from the transformed Mach I cell plates | ||
| + | Grew very slowly and left to grow over night | ||
| + | |||
| + | Designed primers for opening up the pBbE1a backbone for insertion of coil- coil proteins | ||
| + | Coil parts have homology with the flag tag? | ||
| + | |||
| + | Received sequencing results → Results looked good | ||
Latest revision as of 20:30, 17 October 2014
Sequencing results were not good The plasmid did not have the insert
Transformed Gibson product (pBbE1a- CsgA- Spy Tag) into Mach I cells ( attempt 2) Tried varying forms of Gibson product 2 transformations add 2 microliters of Gibson product (1 & 3) 2 transformations adding 4 microliters of Gibson product (2 & 4) Used different transformation tubes → green tops Transformation 1 was probably not good because I was not on the correct setting Transformations 2 and 3 arc’ed first Tried to send pulse again for 2 ---> a good read came up on the equipment Tried to send pulse again for 3----> arc’ed once again Transformation 4 went through without arcing
Grew up another colony from the transformed Mach I cell plates Grew very slowly and left to grow over night
Designed primers for opening up the pBbE1a backbone for insertion of coil- coil proteins Coil parts have homology with the flag tag?
Received sequencing results → Results looked good
