Talk:Team:IvyTech-South Bend/1 September 2010

From 2010.igem.org

9/1/10

Today the team will be making up the same plates as I made yesterday. Professor T. wants new Amp stock made to document how much is used into the stock. Fisher Bio Reagents Ampicillin Sodium Salt Biotech grade. Date Recd 3/18/10 Bp 176-25

1) I have measured out .1287 g Ampicillin stock powder to make a solution of 100 g/mL stock

2) .1287 * .845 = .1087 mg of Active ingedent and dissolving in 1.09 mL H20 with my BR-1000 micropipettor I used the 1000 mark

3) and turned to 009 for the .09 mL added to powder spl. and dissolved

4) using the same pipettor I transferred inside biofume hood 250 mL into 4 sterile ependorph tubes labeled and placed into -80 freezer box.

5) I have poured 4 plates to replace plates from yesterday due to botched Amp

6) Plate 1 – no Amp and 15 mL LB/Agar Plate 2 – 15 mL Amp with 15 mL LB Agar Plate 3 – 30 mL Amp 15 mL Agar/LB Plate 4 – 60 mL Amp 15 mL LB Agar all plates and amp were poured and mixed sterile in Bio-hood I have labeled 1-4 on plates. The Agar was microwaved on beverage setting 4 oz. level four 4 times not allowing to boil over then placed into 40 C Hot water bath to cool amp is added directly into 50 cc sterile tube then added 15 mL Agar to that tube.

Allowed to cool in hood. Now we will streak electro. comp cells on one side and transformed DNA/cells on the other. Electro comp cells I woke up cells into 3 mL of LB Broth I added 2 mL LB/Broth then added electro comp cells to that flicked to mix then added 1 mL more then let sit. Transformation was made by Prof. T.

Streaking

1) Set up Station Cleaning Table with disinfectant along with pipettors and all tools striker, Bunsen Burner

2) Wearing a face covering mask I streaked all 4 plates and placed into incubator overnight. 9/1/10 J.H.