IGEM:MIT/2006/Notebook/2006-6-12

T-Shirt

WE NEED A T-SHIRT DESIGN!

06/12/06 Repeat (from 6/09/06) Transformation Lab

Sources

• Note, used 1/2 of designated source

1. SAMT in PET28 1.6 μg
2. BAMT in PET28 2.0 μg
3. BSMT in PET28 2.0 μg

Dehydrated DNA Recovery (from paper)

• Final DNA Concentration: 1ng/μL (room for better quantification)

1. Add appropriate amount of TE buffer to 1.7mL Eppendorf tubes (500μL for BAMT and BSMT; 400μL for SAMT).
2. Carefully cut out paper that contains DNA and place into Eppen.
   • Ensure submergence with toothpick
   • Wait for 10 minutes

Transformation

1. Followed chemical transformation protocol listed here.
   • Cell Types Transformed:
     • DH5&alpha (big clear tubes with green tape);, Top 10 (Invitrogen) (purple caps), Top 10 (Tom) (big clear tubes), BL21 (Invitrogen) (greenish brown caps)
   • Used 2μL of plasmid/DNA.
   • Heat shock the BL21(DL3) cells
     • 42 c for 50 seconds
   • Used 2xYT
   • Plated 200μL cells

Notes

1. Invitrogen Top 10 BSMT tube fell. It was put on the microfuge for a second to get the cells off the side of the tube.
2. For TOP10 cells use this transformation protocol, which is similar but slightly different from the what we did previously