Lab Notebook

From 2007.igem.org

Revision as of 22:49, 28 August 2007

August 27, 2007

Start Time: 11:00 pm – End Time: 3:00 pm

Members Present: Yusuf, Mimi, Talal

• Carried out plasmid digest (length checks) on I 13507 B and J 37033 B

• Length Check results:

I 13507 B – Overall 3 bands were seen for this plasmid

6-7 (Closer to 7 than to 6)

8-9 (Closer to 9 than to 8)

11-12 (Closer to 11 than to 12)

The number of parts and parts and plasmid match up closely but the first band between 6-7 could be of the uncut plasmid

J 37033 B – Overall 2 bands were seen

8-9 (Closer to 9 than to 8)

10-11 (In the middle)

The bands formed doesn’t match up with the given numbers for its part and plasmid from the registry

• Did Mini-prep overnight for F 1610 with 4 colonies labeled A', B', C', D' and put it in the incubator

August 24, 2007

Time: 5 pm

F/T: Anam

P/T: Talal

-Record the results of the gel Yusuf had made and run (included in table below)

-PD length check for J23100 colony C successful.

-1 band between 6 and 7 on gel

August 23, 2007

Start time: 6:30pm

F/T: Anam

P/T: Fareeha

-Done length check PD for all of A, green circle B, red circle B, black dot B

-Enzymes X and P used for all except for J23100 because the part is too small and so the plasmid that contained it was linearized using enzyme P.

-Buffer 2 was used.

Results:

-All were correct length except for samples from colonies A and B of F1610 and colony A of J37033.

- S01003 (colony A), S01414 (colony B), I13507 (colony A) had 3 bands.

- The 3rd band might have shown up because some of the plasmid was only cut in one location.

To do list:

1) Do a PD length check for the rest of the samples

2) Discuss with Yusuf how much information needs to be recorded when observing the bands in a gel for PD length check

Note: There is one gel left in the fridge. It was made today.

August 22, 2007

Start Time: 5:30 pm – End Time: 7:30 pm Members Present: Yusuf, Anam

• We did Mini Prep overnight and since I did 7 transformations, we decided to do 4 copies of each labeled A,B,C,D so that we have more flexibility in choosing our parts properly. • These are the following symbols we used to clarify all the tubes with the parts

Black circle - I 13507 (AMP Res) (A – D)

Red circle - F 1610 (AMP & KAN Res) (A – D)

Green Dot - S 01640 (AMP Res) (A – D)

Black dot - S 01414 (AMP Res) (A – D)

Red dot - J 37033 (AMP Res) (A – D)

Black line - J 23100 which is a promoter (AMP Res) (A – D)

Red line - S 01003 (AMP Res) (A – D)

• We also labeled our colonies from the Agar plates which were chosen for miniprep overnight as it was instructed by Seema to both Anam and Esther. • The cells which had the promoter J 23100 were light pink colored if Charles and Andy would like to shed some light upon that that would be great.

August 21, 2007

Start Time: 11:00 am

• Charles showed me how to use the registry properly so if anyone needs to know how to use the registry properly just let me know.

o We searched for super parts which is more like a couple of parts ligated together so that it makes our job a lot easier

o So we chose 7 parts to be transformed from the neural network MODIFIED model

• Part timer Muhammad Talal Latif showed up and I instructed him on how to do transformation properly

• So I used 20 l of TE buffer to extract:

o J 37033: 8F, Plate:3 (Amp Res)

o S 01640: 3L, Plate:3 (Amp Res)

• Both of them were from the iGEM 2007 Kit Plates

• We stored the plasmids in the –20C freezer labeled in red DNA remaining

• Added 4 l of J37033 and S01640 to separate competent cell (DH5Z1) eppendorf tubes from the –80C freezer

• So now we are following the Transformation steps from the guidelines posted

• We used two AMP plates made from before

• We also made 8 AMP plates where 4 is going to be used today for the other four parts which we also transformed:

o I 13507: (Used form 2005 stock) (AMP Res)

o S 01003: 20O, Plate:1 (AMP Res)

o J 23100: 21E, Plate: 3 (AMP Res)

o S 01414: 22O, Plate: 1 (AMP Res)

• We also made 3 AMP and KAN plates where 1 is going to be used:

o F 1610: 1B, Plate: 2, (AMP and KAN Resistant)

• CONFUSION: While making the AMP and KAN plates we used AMP of 100mg/ml whereas KAN was 30mg/ml. So even though KAN was of different concentration we used the same quantity.

o CHARLES AND ANDY GIVE US SOME INSIGHT ON TO THIS TOPIC

• So we left all 7 plates for overnight incubation and hopefully we see some results tomorrow

• Everybody please welcome SeungMe (part time) who came in and helped me finish the work today and of course Charles