Team:Shanghai City/Notebook
Notebook
Hi! Welcome to the notebook section! Here I will briefly introduce what are included in our notebooks.
There are three types of things are included in our notebooks: Systems, Calculations(&Error), and Background knowledge.
--Systems:
Formulas are the most important part in our experiments. No matter whether you have some background information about our experiments, if you can follow the formulas and instructions in our notebooks, than everyone can do the experiments. These formula not only record function and purpose, but also materials that experiment need and amount of each materials need. Our formulas recorded in our notebooks included:
1.25μl system for PCR
2.20μl system for Cas12a reaction(Differ with different primers)
3.20μl system for crRNA transcription
4.50μl system for Plasmid DNA transformation(Differ with different templates)
5.Template transformation instructions through E.coli
6.Several 25μl systems for PCR with different amounts of variables
7.Several 20μl systems for Cas12a reaction with different amounts of variables
Here are some examples of systems:
--Calculations(&Error):
Calculations part includes some calculation we made during dilute the crRNA, templates, and primers. It also involves several formulas for different amounts of variables which got from the wrong results in previous experiments. Contain come error analysis.
Here are some examples of Calculations(&Error):
Here are category of each notes based on type of experiment we did that follow the sequence of event:
#1:
In these two notes, we did the crRNA transcription to made the crRNA that we need in further experiments. These crRNA will be used with cas12a protein to find out true primer in our mixed primer which contains true primer and false primer. Using the T7 promoter, nTP and several templates() that can transcript different crRNA.
#2:
Before we did system of cas12a reaction, we wanted to test the result of mixed primer which contains false and true primer, and this was kind of control group in our experiment to compare the result when we went through the cas12a reaction and got the theoretically true primer. In this page of note, we also measured the needing of different temperature for later cas12a reaction experiment.
#3:
After we did the control group, we began to test encryption of mixed primer through cas12a. Our purpose in this part was to find whether using cas12a reaction we can get true primer in order to solve true message that hided in mixed DNA template. Here has two system, one is cas12a reaction system and another one is PCR system. We struggled for long time in this testing because the results were always unstable and vague, and not strong enough to support the finding of message, so we tried many different amounts of materials need, mainly change in amount of cas12a protein and primer, and many different values of variables such as reaction temperature and time. There were many groups in these notes record the changing of amount and values.
#4:
Based on the error we often occurred in testing cas12a reaction, we made error analysis and finally determine two possible error: primer was not pure enough or primer specify. By this way, our AI helped us to find a way solve these problems through introduce us about plasmid DNA and E.coli which can not only copy the DNA template but also can help to purify the primer.
#5:
This note contains procedure about how can we do primer purification and DNA transformation. Through adding E.coli, plasmid DNA, our DNA template, and changing of temperature, we can produce the E.coli we need and foster them and finally get purified template.
Thus, the notebook part finished, and these notes were not present all of our experiments and there were much more detailed information that not recorded in our notes but contains in our experiment and results.