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Team:Colombia/Journal
From 2014.igem.org
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=June 24= | =June 24= | ||
| - | Today was the first day at the lab!! Finally! After | + | Today was the first day at the lab!! Finally! After finding out the parts we ordered from the registry had been sitting in a locker for the last two weeks and managing to salvage them we re finally up and running. First thing first, today we are making electrocompetent cells and transforming our first parts. We are using TOP 10 strain to make our transformations. We are transforming mRFP (BBa_K518012), pBAD (BBa_K206000), tetR inverter (BBa_Q04400), amilCp (BBa_K592025), pBAD-CqsA (BBa_K581011), pTet-LuxOD47E (BBa_K218017), pqrr4-GFP (BBa_K581009) and LuxU-CqsS-LuxO (BBa_K581010). We are starting with the Interlab project, so we are transforming those parts as well. |
| + | |||
| + | We just realized we are all out of our kanamycin stock. We need to prepare some more tomorrow. | ||
=June 25= | =June 25= | ||
| + | |||
| + | We have transformants! Well, almost. Nothing grew on the IL4 plate :(. But nothing to worry about, we are just repeating the transformations. We made passes to liquid media of single colonies of all the other parts and we are hoping to do miniprep of all them tomorrow. | ||
| + | |||
| + | We also cryopreserved all the bacteria we got from the registry. Just made a fresh new Kanamycin stock, so we can now transform tetR inverter. | ||
| + | |||
| + | =June 26= | ||
| + | |||
| + | Our first minipreps were unsuccessful and nothing grew on the tetR inverter plate. | ||
| + | |||
| + | We just realized the tetR inverter part has two other twins in the registry. We are transforming those as well and hope one of them works. | ||
| + | |||
| + | =June 27= | ||
| + | |||
| + | Today we prepared homemade lysis buffer for our minipreps, but apparently it wasn't very effective. We could not see any DNA after running our miniprerps in a gel. | ||
| + | |||
| + | =July 1= | ||
| + | |||
| + | After a long weekend, we are back in business. We have colonies in all the plates with the trnasformations of the tetR inverter! We made passes of all three parts to liquid LB for miniprep tomorrow. | ||
| + | |||
| + | =July 2= | ||
| + | |||
| + | We repeated the miniprep of all the parts and finally got it right this time! All molecular weights correspond to what we expected! | ||
| + | |||
| + | =July 3= | ||
| + | |||
| + | Now that we have all the parts, we can start building our construct! We are starting with the Interlab parts for now. Today we digested both promoters and GFP , ligated them and transformed them using our electrocompetent TOP 10 E. coli. | ||
| + | |||
| + | =July 4= | ||
| + | All our transformations were unsuccessful. After running the digestions in a gel we realized that the digestion was not complete. We did some positive controls on all the enzymes to verify that they are working properly, but it all seems to be working fine. | ||
| + | |||
| + | =July 7= | ||
| + | |||
| + | Today we are measuring fluorescence of the first part of the Interlab Study! We are also trying to begin joining the first two parts of our construct tetR inverter and mRFP. Here is some doodles we used to explain to the new iGemers how the process of digestion and ligation works and the logic behind it. | ||
Revision as of 16:39, 15 October 2014
Contents |
June 24
Today was the first day at the lab!! Finally! After finding out the parts we ordered from the registry had been sitting in a locker for the last two weeks and managing to salvage them we re finally up and running. First thing first, today we are making electrocompetent cells and transforming our first parts. We are using TOP 10 strain to make our transformations. We are transforming mRFP (BBa_K518012), pBAD (BBa_K206000), tetR inverter (BBa_Q04400), amilCp (BBa_K592025), pBAD-CqsA (BBa_K581011), pTet-LuxOD47E (BBa_K218017), pqrr4-GFP (BBa_K581009) and LuxU-CqsS-LuxO (BBa_K581010). We are starting with the Interlab project, so we are transforming those parts as well.
We just realized we are all out of our kanamycin stock. We need to prepare some more tomorrow.
June 25
We have transformants! Well, almost. Nothing grew on the IL4 plate :(. But nothing to worry about, we are just repeating the transformations. We made passes to liquid media of single colonies of all the other parts and we are hoping to do miniprep of all them tomorrow.
We also cryopreserved all the bacteria we got from the registry. Just made a fresh new Kanamycin stock, so we can now transform tetR inverter.
June 26
Our first minipreps were unsuccessful and nothing grew on the tetR inverter plate.
We just realized the tetR inverter part has two other twins in the registry. We are transforming those as well and hope one of them works.
June 27
Today we prepared homemade lysis buffer for our minipreps, but apparently it wasn't very effective. We could not see any DNA after running our miniprerps in a gel.
July 1
After a long weekend, we are back in business. We have colonies in all the plates with the trnasformations of the tetR inverter! We made passes of all three parts to liquid LB for miniprep tomorrow.
July 2
We repeated the miniprep of all the parts and finally got it right this time! All molecular weights correspond to what we expected!
July 3
Now that we have all the parts, we can start building our construct! We are starting with the Interlab parts for now. Today we digested both promoters and GFP , ligated them and transformed them using our electrocompetent TOP 10 E. coli.
July 4
All our transformations were unsuccessful. After running the digestions in a gel we realized that the digestion was not complete. We did some positive controls on all the enzymes to verify that they are working properly, but it all seems to be working fine.
