Since then we remade both our CARs and have gotten things to work pretty well
We have nearly finished every part and are now well into device making… for the Meso car that is.
In the assay our NKL -GFP / K562-CD19 and NKL-anti-CD / K562- CD19
SHOWED THE SAME KILLING RATE; the car was a failure in our assay, but since it worked for the people we got it from, it is very odd
The super buddies are trouble shooting the issue.
Ray and heather have left us
they were loads of fun
Ray had a good sense of humor and Heather was always enthusiastic
Ray still owes me some surf n turf
HELL YEAH
(probably not tho)
school starts in a weeeek
i dont wanna go back
iGEM is so much fun
i wanna go to college just like the other people here
>:(
(oh well)
BLOG BLOG BLOG
I went camping over the weekend
i was the only human to go swimming in roundtop lake and that other lake up above woods lake.
There were a bunch of golden retrievers there though,
Sam the man 1 Sam the golden retriever 0
( fetch in the water i won)
now back at working
going yum cha with friends for long lunch
*deliberate nose intake
YUP
-sam
Sam didn’t log out of his blog. He is lucky someone lacking bad intentions found his blog. Sam should learn to log out of his blog. :)
so after a bout a month of hard work generating “parts” or desired sequences of DNA that code for proteins with restriction sites that we want, Connor and I finally finished our AB parts, the two CARs ( Chimeric Antigen Receptors) which are some of the most vital parts in our project. However, our happiness was short lived as our work was ripped from our hands and shatter into a million pieces. The culprit? Middle management.
In the initial stages of generating parts, we were given templates to follow which worked out pretty well. In the next week we had to scrap all our parts and use new templates supplied by Jason to use. SO after about three weeks of working on my AB part i finally finished it monday and digested it successfully tuesday. All was good in the world and our deadline was gonna be made. Then yesterday James, our PI, looks into our template and realizes that he does like the template for our AB parts,
the regular kozak seq is GCC GCC ACC then gene start….
the one Jason had us use was GCC GCC ACC ATG GGA gene start.
because these parts are receptor proteins, James is unsure if this will make it not work
the Assay is $700, we run every assay in parallel. Maybe isn’t worth a 1500 test.
we start again monday
at best a set back of a week
RAGE
so
in the morning before most people come to work the main bathroom lights are on motion sensors as to not waste energy. But something cool i found is that if you walk very slowly, the sensor doesnt pick you up and the main lights stay off
so i was all like
walkin hella slowly into the bathroom, and i snuck almost all the way up to the urnal
when suddenly a guy walked into the bathroom and the lights turned on
spoiled my ninja fun »>:(
in other news….. [ aka the related matters]
i finally got a PCR to work, but got bad yeild like 3.5 ng
as apposed to like the 708ng that i once got and my TOPO reaction failed so i had to restart all over again
^ happened at 7pm so the whole day was wasted
but since it was friday everyone was on happy hour==
5—>6 free beer soda and chips
so i got some chips and hansens orange soda
:)
good day
when i got to work Jacinto let me play whit liquid nitrogen. I froze cell dishes and tubes and smashed them and it was cool. i also froze tape and shattered it and it was coooooooool.
Bad day:
my sequencing results came in. 811 -> 815 M13F failed. and my P85 samples all had weird results that warrented restarting everything.
My PCr failed because i ran out of Phusion the first time 2 hours down the drain
my 2nd PCR failed for no reason wasted 4 hours
i accidently left Phusion polymerase on the bench -$400 =[
my 3rd PCR reaction failed wasted 6 hours
but then i had a good day after work
and i wrestled with my friend
WRAAAAAHH PENT UP RAGE FROM FAILURE HEAD KICK HEAD KICK LEFT HOOK
Well days have been a bit monotonous, a few days of miniprepping the same samples of DNA, sending things in sequnecing, and other stuff like that
while its alot of the same work, its still pretty fun to run the experiments the days just fly by right now
well its been a very long time since i have posted something…
today is friday and for the afternoon we are going to berkeley for a BBQ / soccer / fun
with other iGEM teams
today i really messed something up in teh lab but i dont think its gonna be that bad
>.> whew
iGEM is, as ever, so much work, and as much fun as the paper clip [ so much fun]
worked a day from 740am to 930pm and i was tired that i just died after that
i think that we as a group are working more smoothly than before
things are going great and its lots of fun
science is science
life life
and iGEM kicks ass
sam out
Today we started our first wetlabs, by this i mean that for once we actually did shit.
In the morning ryan and eric had us inoculate colonies ( grow liquid culture) for the purpose of running a miniprep, but that didnt happen so our cells are still growing overnight
we also got to run our first PCR today, it was from the primers we ordered.
it feels like all that slave labor with the computers payed off :)
on a side note, we had a QB3 pizza meeting. QB3 is like a science collaboration around the bay area and stuff,
they fed us weird pizza
and the speaker talked bout stuff that made me say “gattaca” to john knowingly, and a little afraid too
today is monday, the start of week three of iGEM
bootcamp ended last week and i left you with vivid memories of struggle, love, deciet, valor, glory, and romance… or something like that.
Today i tell you that work insofar is slow, we are in the early phases of figuring out what we need to work with and how to get it. We are left to fend for ourselves for the most part, with help from our buddies. Lianna and i [yes…. yet again…] have gotten Russell as our buddy. He is pretty much the most awesome guy. He is an early starter and early leaver, and frequently comes in to check on us. He is soft spoken and helpful, a regular gentlemen if it does ya
i did work on the same sequece a few times, trying to get the hang of how it all works. orientation of DNA and such is pretty difficult, but once you get it you got it
i hope =[
good times are on their way though, lab work will being soon and life will be lost in the myriad of mini preps and experiments.
well CR its been nearly two weeks since i started iGEM
i was assigned to Team A consisting of Heather, Crystal, Lianna [ as always], connor, and me
we were assigned to work on the cytotoxic payload of NK cells for our team challenge project.
We dont work very well as a team, but it all worked out i guess.
[crystal is a perfectionist
connor usually couldnt care less
im lazy
and lianna is hungry]
but like I said, somehow we made it work.
iGEM is a whole ton of fun. I work best under pressure, so the team challenge was probably more fun for me than for most people. I love to find innovative solutions to problems, even when people like Wendel Lim around to drill you on all those fine point you just dont quite know.
My idea was to link Granzymes together for a more effective, which grew into our main functional idea, i hope it works out, itll be a fun project to experiment with
We have great time on our own and all together. I swear you wouldnt think we were doing anything at all if u werent looking at our computers
time flies too fast, but every second is a blast.
sam out
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