Team:Calgary/22 July 2009

July M T W T F S S     1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

Carol
Chinmoyee
Emily
Fahd
Iman
Jamie
Jeremy
Katie
Kevin
Mandy
Patrick
Prima
Stefan
Vicki

May M T W T F S S         1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

June M T W T F S S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

July M T W T F S S     1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

August M T W T F S S           1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

September M T W T F S S   1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

October M T W T F S S       1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

• Prepared sterilized LB broth for growing cells. Followed procedure found in the protocol page for making competent cells.
• After completion of making competent cells, transformed ligated products (pCS26+promoters ligated using NEB Quick Ligase) into XL Gold Ultracompetent cells. These cells were plated on LB+Kan agar plates and was incubated overnight at 37^oC.
• As well, as a control, we transformed pBluescript into both XL Gold Ultracompetent cells and Top 10 Cells and plated cells on LB plates to incubate overnight at 37^oC.

WIKI CODING HERE

• Did a miniprep of overnight cultures and took concentrations.
• Set up a Restriction Digest for verification (that the B0015 terminator is present in my contruct) with XbaI and PstI. If successful, we expected to see bands at about 1.7 kb and 3 kb for the contruct (J13002-LuxOD47E-B0015) and the psb1ac3 vector respectively.

Lane 1- J13002-LuxOD47E-B0015 C5, cut with XbaI and PstI
Lane 2- C5 uncut
Lane 3- C6 cut
Lane 4- C6 uncut
Lane 5- C7 cut
Lane 6- C7 uncut
Lane 7- C8 cut
Lane 8- C8 uncut
Lane 9- Size control- J13002-LuxOD47E
Lane 10- Size control- BBK LuxOD47E

• Analysis: From this gel it looks like colonies 5, 6 and 7 may have worked (contain the B0015 terminator) as we see two bands at the expected sizes. Colony 8 has some random bands, so I will ignore this one. Colony 6 looks the most promising as it is the cleanest (no random bands). I will proceed by sending this colony down for sequencing.

• Wrote out a lab blog for this week that I shall post tomorrow morning.

Today, I concentrated my energies at marketing our iGEM project and looking at the ethical aspect of our project. Here is what I did today:

1) I was very happy when i read NEXEN INC.’s e-mail that they would give us 3000 dollars. Nexen Inc. would be our first silver sponsor.

2) I e-mailed a thank you letter to Nexen Inc. and also sent them PDF copies of our Sponsor webpage. I will also mail in a thank you letter.

3) Helped approve/edit the July's newletter.

4) E-mailed the lab equipment and reagents list to VWR

5) Called Life Technologies and left her a voicemail.

6) Called EMD biochemicals and left a voicemail for us.

7) Called Bietz Resources Ltd. He will not be able to help us but he has offered his services(contacts) to our team.

8) Called Albian Sands Energy Ltd. and left a voicemail for him

9) Called Alberta Pacific Forest Industries Inc. and left a voicemail for him.

10) Helped out in the lab: I was putting tubes in the racks and closing the lids.

WIKI CODING HERE

WIKI CODING HERE

WIKI CODING HERE

The bacterial transformation activity was fixed in the morning and I added to the conditionals that tell you if the question has already been answered so once a question has been answered it cannot be answered again unless the quiz has been completed or reset at any time. I also added a further instructions option to the PCR machine, which gives a notecard for now, but I am interested in trying the instructions as audio. The instructions are for those who throw themselves into the lab first thing or those who need a reminder of how to operate their tools in second life.

The first and second construction site now use the notecard reader and I added more options to the Sequencing station so more can be sequenced. Now I just need to know the sequence of base pairs for each notecard I will be using. At the moment the notecards I am testing with contain random data. Ligating two genes together may be something I will be adding to the restriction digest activity once I can get it all working together since it is just adding more conditions. However, I would have to redesign how you could put things together so I believe I will hold off coding anymore of this until the activity has been finished. Tomorrow morning I can quickly complete the third construction site script and test with generic notecards.

One thing I'm not really comfortable with the dataserver event yet, which is very important to reading the notecard so I think it would be useful for me to trace the code within this event and determine exactly what it does just so I completely understand how notecard reading works. Since we should probably get started on the base of the spiral that will contain levels I have started to write up some more notecards that are even more basic than the ones I have been writing (transcription, translation etc.) like:

• What is a gene?
• What is DNA? etc.

I have sequenced by Colony 1 and 9 of my Pqrr4+Boo34 (RBS) because they were the closest to the positive controls and the cleanest lanes. For this, I have only used Biobrick CP forward sequencing primers, and no reverse. I only used forward and not reverse because the piece is quite short, and forward primer is enough to read every base pair. These are hopefully being sequenced right now and I am waiting for the result to be in.

Two types of competent cells were made today, and they were TOP10 and XL Gold. Top 10 cells were made because we were running out on them, and XL gold was made because Top10 seemed to be not competent enough for Carol’s luciferase.

WIKI CODING HERE

WIKI CODING HERE

Today:

I followed up with: Company 1: called Dr. Maloney’s asst. and she said he’s unable to meet with us this month but she’ll discuss our sponsorship pkg with him. I was thinking of calling his assistant again tomorrow. Then I’ll follow up on the sponsorship pkg on July 208h

Company 2 Currently looking over our sponsorship pkg. I called and the Marketing In-charge was not in the office so I left a msg: follow up July 23 (tomorrow). Everytime I leave a, I’m saying that we’re not asking for large grants and that our donations have ranged from 200 to 6000 dollars so any contribution will be greatly appreciated by the team. I told the rest of the marketing team to say the same thing.

Company 3 -called to follow up- the Service Desk lady said she forwarded our pkg to the right person and since they didn’t reply back it probably means they’re not interested. I asked for that person’s contact info, but she said it’s against their policy to give out staff contact info.

Company 4 -currently looking at our sponsorship pkg – called but she wasn’t in the office, left a msg so follow up July 23

Company 5 -called, not in the office, so I left a msg. Follow up July 23.

Company 6 -called the lady for the 20th time in the last 10 days, she’s not in her office and there’s APPRENTLY no one else who handles sponsorship. GRRRR. But I’m gonna keep harassing her. Customer service APPRENTLY has no way to get a hold of her but they saw coming in and out of her office so she’s not on vacation.

GlaxoSmithKline - this is the new company that I found yesterday. Called, left a voice mail. I need her email address so as soon as I get a hold of her I’ll talk to her and send pkg

I also edited the newsletter that Jamie sent to the marketing team last night. I’ll discuss the corrections tomorrow. Then I checked over a couple of marketing emails that Jeremy and Fahd were sending out companies. Then Fahd and I wrote up an email toVWR and Mandel Scientific requesting specifically the reagents and pipets that Thane wanted us to order. We checked the email over with Jeremy before sending.

For about 20 mins, I was pointing out a few things/suggestions for the wiki with Mandy.

I was working in Second Life for most of today, specifically the eukaryotic cell. I scrapped the previous endoplasmic reticulum and installed a new one and it looks much better. Scripts were added to make it "sparkle" in order to give it that ribosomal look. Smooth ER was also created today. Experimentation with particle systems proved to be a worthwhile endeavor. I had trouble with changing the settings, but I eventually figured out how. the result was vesicles coming out of the Golgi body periodically (you should check it out!). I have also chosen a location for the cell and thus, was able to script some of the organelles to move and expand. Tomorrow there will be a meeting with Gregor about ethics at 10 so that should be interesting.

I wrote a preliminary introduction and most of the materials and methods section for the LuxO D47A mutant.