Lab Notebook
From 2007.igem.org
Contents |
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 (DH5Z1) 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
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 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 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 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
