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From 2010.igem.org

Protocols

Dip and swirl cloning

This protocol is essentially the same as the Standard BioBrick Cloning protocol used in our lab with the exception of using a minimal amount of restriction enzymes.

• Restriction Digest

1. 1. Add 0.3 μl of 10X NEBbuffer 2
   2. Add 0.3 μl 0f 10X BSA
   3. Add 30 ng of DNA (plasmid or PCR product)
   4. Dip the pipette tip in the enzyme solution and move it around (swirl). Place the tip in your reaction mixture, move it around (swirl) again, and rinse it.
   5. Add water to a final volume of 3 μl.

• The remaining steps of the protocol are identical to that of the Standard BioBrick Cloning protocol.

N.B. This protocol is particularly suitable for when the enzyme solution has almost run out, but you still need to perform a number of digestions. This protocol is longer than the Standard BioBrick Cloning protocol. In addition, if the level of the enzyme solution is high, the capillarity action causes more than enough of the enzyme to enter the tip, hence, rendering this approach pointless.