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Team:Tsinghua-A/Notebook
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Revision as of 15:08, 5 October 2011
Notebook Section
07.14.
Disturbed but happy
Today, the summer semester ends and the biological experiment comes. As we are green-
hands in the field of biology, everyone feels very disturbed. Doctor Chen encourages us to cheer
up and teaches us how to transform. After one and a half hour, we finally get it. We successfully
finish the transformation of B0034!
07.15.
Sad but expecting
Today, we continue to do cell transformation. We get our medium of B0034 from incubator.
Unfortunately, we find that there is nothing in the medium. I am very sad, but our captain
encourages us to try again. We manage to do it with the rest B0034 in the kit. After one hour, the
assignment today is completed. We wish that our effort wouldn’t be in vain.
07.16.
Excited
We get it! The transformation of B0034 proves to be a success. Everyone here is excited and
ready to finish the transformation of C0079, B0015, R0079, J23116 and J37034.At the same
time, we contact Doctor Chen and tell him the exciting news. He congratulates us and promise to
teach us the procedure of shaking bacteria.
07.17.
Stirring
Today, we learn how to pick the right colonies that we need and the protocol of shaking
bacteria. After some practice, we learn the point of this step, and quickly finish our work
(including B0034, C0079, B0015, R0079, J23116 and J37034) today.
07.18.
Damn difficult!
Today, Doctor Chen teaches us the protocol of plasmid extraction. Compared with the
experiments we have done before, plasmid extraction is really a big problem. To finish plasmid
extraction, there are eleven steps that we have to do. The work takes us more than one and a
half hours. With the help of Doctor Chen, we finally finish the extraction of
B0034,C0079,B0015,R0079,J23116 and J37034.
07.19.
That’s easy!
Today, we finish the transformation of K145270,R0062,P0440 and E0420.
07.20.
In low spirit
Unluckily, we find that there is nothing in the medium of P0440 and R0062, but the
transformation of K145270 and E0420 proves to be a success. We have a discussion about the
protocol of transformation. After 10 minutes, we doubt that the failure of the transformation
results in the incorrect operating of DH5. Then we carefully do the transformation of P0440 and
R0062.
07.21.
Happy
The effort we took yesterday is not in vain. We successfully complete the transformation of
P0440 and R0062. Then we begin to pick the right colonies from the medium. After that, we put
all the tubes into the shaker and start to shake the bacteria.07.22
Not easy
It is an awful day today, because we have to do the plasmid extraction of K145270, R0062,
P0440 and E0420. Without the help of Doctor Chen, the steps of plasmid extraction are too hard
for us to complete them without any mistakes. We waste many centrifuge tubes and pipet tips.
However, we finally make it.
07.24.
Lazy
After our weekends, we come back to our lab. The task today is to finish the transformation
of R0040, K081016.
07.25.
Lazy again
Today, we complete the transformation of I0500, K081009, E0430.
07.27.
Get confused
Today, we are surprised to find that the transformation of I0500 fails. We try to find the
reason. After about 15 minutes, we don’t find any mistakes in our procedure. As a result, we
contact Doctor Chen. He promises us that he will come tomorrow.
07.28.
Rewarding
Today, we have a discussion about the failure of the transformation. Doctor Chen supposes
that there is some problem with the Kan medium. So he writes down the medium formula and
asks us to make up some Kan mediums. With his help, we learn the method to use the electronic
balance. Finally we use our new Kan mediums to do the transformation of I0500.
07.29.
Damn!
We are very disappointed when we take out our mediums. The transformation fails again.
Then we decide to find some information from the Internet. Maybe other IGEM teams face the
same problem.
08.01.
Terrible day
After our weekend,we exchange our findings that searched from the Internet.We are very
angry when we find that the plasmid doesn’t work.Then,we tell the message to Doctor Chen and
begin to find another plasmid to replace I0500.
08.02.
We make it!
Everyone works hard to find the new plasmid. Where there's a will, there's a way. We
successfully outcrop a new plasmid named I13453 which proves to be a workable plasmid.
Then we quickly complete the transformation of it.
08.03.
Nope, it's Chuck Testa!
We succeed to do the transformation of I13453.And then we pick the right colonies from the
medium (including K081009,E0430 and I13453), and put the tubes into shaker.
08.04.
Another easy day. Today, our task is to finish the plasmid extraction of K081009, E0430 and I13453. After the
practice we have done before, we find that it is not too difficult to make it. We only spend one
hour extracting the plasmid.
08.08.
New protocol
Today, Ms. Yang teaches us the protocol of agarose gel electrophoresis. Compared with
plasmid extraction, electrophoresis is easier. But we need to make the gel and grasp the method
to use microwave oven.
08.09.
A blue day.
Today, we do the agarose gel electrophoresis of B0034, C0079, B0015, R0079, J23116 and
J37034. Unfortunately, we find that our plasmid extraction of C0079, R0079 and J23116 fails. It
proves that our chosen colonies are not target bacteria colonies. Therefore, we take out our
medium and repick the other colonies and put the tubes into shaker.
08.10.
As usual.
Today, we complete the plasmid extraction of C0079, R0079, J23116.
08.11.
More than exciting!
The plasmid extraction is successful. We observe the plasmid in the agarose gel
(including C0079, R0079, J23116). And then we do the agarose gel electrophoresis of K145270,
R0062, P0440 and E0420. We find that we have to redo the plasmid extraction of K145270,
R0062. Therefore, we take out our medium and repick the other colonies and put the tubes into
shaker.
08.12.
Just a day.
Today, we complete the plasmid extraction of K145270 and R0062.
08.20.
Blissful!
The plasmid extraction is successful. We observe the plasmid in the agarose gel
(including C0079, R0079, J23116).But the plasmid strips are very strange. So we do the
electrophoresis of C0079, R0079, J23116, K081009, E0430 and I13453.We find that we have to
redo the plasmid extraction of K081009,C0079. Therefore, we take out our medium and repick
the other colonies and put the tubes into shaker.
08.21.
Nothing special.
Today, we complete the plasmid extraction of K081009,C0079.
08.27.
cheerful
The plasmid extraction is successful. All the plasmid we need have already
been extracted. Then Ms. Yang shows us the method of digestion. At first, we learn the use of
Bio Photometer. And then we need calculate the mole of the target plasmid. At last, we put the
solution to target plasmid, corresponding digestion enzyme, double distilled water and BSA into a
tube (including B0034, C0079, B0015).
08.28
.A tragedy
Unfortunately, we find that our digestion fails after the agarose gel electrophoresis. So we
have to redo the digestion. Luckily, we make it and get our first gene--------B0034,C0079,B0015.
09.03.
Ongoing
Today, we do the digestion of R0079, J23116, J37034.
09.04.
Ongoing
Today, we do the digestion of K145270, R0062, P0440 and E0420.
09.05.
Ongoing
Today, we do the digestion of K081009, E0430 and I13453.
09.06.
Something new
Today, Ms. Yang shows the protocol of connection. Like digestion, the procedure of
connection is not hard. But it takes 16 hours. So we do the connection of K081009,E0430.
09.07.
A hope
We do the transformation of our new plasmid, KE.
09.08.
If we do right?
We pick the right colonies from the medium, and put the tubes into shaker.
09.09.
Desperate
Today, we complete the plasmid extraction of KE. Unfortunately, we find that our connection
fails after the agarose gel electrophoresis. Then we redo the transformation of KE.
09.10.
Fighting
We pick the right colonies from the medium, and put the tubes into shaker.
09.11.
Still fighting
We complete the plasmid extraction of KE. Ya-da! Our connection success! We got our first
part, KE!
09.13.
Still fighting
We do the connection of R0040 and K081016, R0062 and P0440, R0062 and E0420.
09.14.
Still fighting
We do the transformation of our new plasmid-------RK, RP, RE.
09.15.
Still fighting
We pick the right colonies from the medium, and put the tubes into shaker.09.16.
Still fighting
We complete the plasmid extraction of RK, RP, RE. The connection of RP and RE success! We
got our next two parts-------RP and RE! However, the connection of RK fails. We have to do it
again .But we believe that our project will be finished soon!
09.17.
No much time left.
We do the connection of B0015 and C0079, R0079 and J37034.
09.18.
Speed up
We do the transformation of our new plasmid-------CB, RJ.
09.19.
As usual
We pick the right colonies from the medium, and put the tubes into shaker (including RK).
09.20.
Sprinting
We complete the plasmid extraction of CB, RJ, RK. Ya-da! The connection of JC and RK
success! We got our next two parts------- CB and RK. But the connection of RJ is not successful.
Thus, we decide to use a plasmid of GFP instead of J37034.
09.21.
Sprinting
We find a new plasmid that can replace the part JC, so we use it instead of our new part JC
and we name it “K”. We do the transformation of “K” and GFP. At the same time, we do the
digestion of RK, RP, I13453, RP, RE, K145270, R0079. After the digestion, we do the connection of
IKE, RPRE, KRP.
09.22.
Sprinting
We pick the right colonies from the medium, and put the tubes into shaker. And we do the
transformation of our new plasmid-------IKE, RPRE, KRP.
09.23.
Sprinting
We pick the right colonies from the medium, and put the tubes into shaker. We complete
the plasmid extraction of “K” and GFP. We do the digestion of “K” and GFP. After that, we do the
connection of “K” and CB, R0079 and GFP.
09.24.
Sprinting
We complete the plasmid extraction of IKE, RPRE, KRP, we get our new parts-------IKE,RPRE
and KRP. Then, we do the digestion of IKE, RPRE, KRP. After that, we do the connection of RKIKE,
KRPRE. At last, we do the transformation of our new plasmid-------KCB,RG.
09.25.
Sprinting
We do the transformation of our new plasmid-------RKIKE, KRPRE. After that, we pick the
right colonies from the medium, and put the tubes into shaker (including KCB and RG).09.26.
Sprinting
We pick the right colonies from the medium, and put the tubes into shaker (including RKIKE,
KRPRE). Then, we do the plasmid extraction of KCB and RG. Fortunately, we succeed. We get our
new parts-------KCB and RG. We do the digestion of KCB, RG. After that, we do the connection of
KCB and RG.
09.27.
Sprinting
We complete the plasmid extraction of RKIKE, KRPRE. we get our new parts-------RKIKE,
KRPRE. We do the transformation of our new plasmid-------KCBRG.
09.28.
Sprinting
We pick the right colonies from the medium, and put the tubes into shaker (including KCBRG
only).
09.29.
Yeah, we make it!
We complete the plasmid extraction of KCBRG, we get our new parts-------KCBRG. Till now,
our project has been completed. We are so excited that we have a big dinner in the evening.
