Minnesota/4 August 2008

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1. Figure out what going to send in products for sequencing
2. Meeting with Yiannis: (1) Make sure powerpoint does not exceed 30 minutes, send it to Yiannis on Wednesday for him to review. (2) Write paper with materials and methods that can point to specific papers that we followed instead of writing out entire procedures. (3) Have iGEM requirements all finished before week ends.
3. Ligations: Ligate the digested products from 08-01-08. Follow the table below:


Parts 10x Buffer H20 prev. lig. + BV DNA Insert DNA T4 DNA Ligase Total
BV + Tet/p22 + RFP + Term 4.0uL 4.0uL 5.0uL 23.0uL 4.0uL 40.0uL
BV + Tet pro + LAMBDAcI + Term 4.0uL 4.0uL 5.0uL 23.0uL 4.0uL 40.0uL
BV + Lac/LAMBDA + GFP 1 4.0uL 4.0uL 5.0uL 23.0uL 4.0uL 40.0uL
BV + Lac/LAMBDA + GFP 2 4.0uL 4.0uL 5.0uL 23.0uL 4.0uL 40.0uL
4. Plate Ligated prodcuts: Plate the ligated products on ampicillin resistant plates. Allow to grow in incubator @ 37C for 8-12 hours.
5. Pick Colonies from plated ligation products: Pick 5 colonies from every one of the 4 plates (making 20 2.0mL tubes) and place 2mL cultures in incubator @ 37C for 12 hours with shaking @ 220rpm's.