1. Kit location
   1. Plate 4, Well 18A

To use the DNA in the Distribution Kit you may follow these instructions:

1. With a pipette tip, punch a hole through the foil cover into the corresponding well of the Biobrick™-standard part that you want.
2. Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes to make sure the dried DNA is fully resuspended.
3. Transform 1 or 2uL of the resuspended DNA into your desired competent cells,

This protocol can be used to transform chemically competent (i.e. from CaCl2) with a miniprepped plasmid or a ligation product.

Note: Never vortex competent cells. Mix cells by gentle shaking.

1. Thaw competent cells on ice. These can be prepared using the CaCl2 protocol.
2. Place 20 ul of cells in a pre-chilled Eppendorf tube.
   • For an Intact Vector: Add 0.5 ul or less to the chilled cells
3. Mix gently by flicking the tube.
4. Chill on ice for 10 minutes. This step is optional, but can improve yields when transforming a ligation product.
5. Heat shock at 42 °C for 30 seconds.
6. Return to ice for 2 minutes.
7. Add 200 ul LB medium and recover the cells by shaking at 37 °C.Kanamycin: 120 minutes
8. Plate out the cells on selective LB.Note: 200 ul is the maximum volume of liquid that an LB plate can absorb.
9. Incubate at 37 °C. Transformants should appear within 12 hrs.

July 5th

Bacterias plated yesterday growed and made big and small colonies