From 2007.igem.org

August 21st, 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 hell 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