Processing Clones from Whatman FTA Discs (From DGRC)

Required: determine what antibiotic -- DGRC recommends 34mg/ml chloramphenicol 1000X stock)
 * 2 LB/antibiotic plates per disc (check resistance gene in plasmid to


 * 1X sterile TE


 * Thawed competent cells on ice* (Do not use XL10GOLD)


 * 500uL SOC medium

* DGRC recommends DH5alpha or equivalent. Cells should have efficiency of 1 x 10^7 cfu/ug or higher. Note that XL10 GOLD cells carry a chloramphenicol-resistance gene on the F’ episome.
 * Sterile microfuge tubes
 * 1) Add 50uL sterile 1X TE to the microfuge tube with the clone disc and pipet up and down quickly twice. Remove the TE immediately and discard. Do not leave the TE on the disc for more than 2 seconds.**
 * 2) Place the tube on ice.
 * 3) Add 50uL competent cells to tube with disc and incubate on ice for 30 minutes, vortexing for 1 second halfway and immediately returning to ice.
 * 4) Heat shock cells/disc for 2 minutes at 37℃.
 * 5) Transfer cells only (leave behind disc) to 500uL SOC medium and incubate with shaking for 2 hours.
 * 6) Plate 100uL on one plate, and the remaining on another plate (should be about 450uL).
 * 7) Incubate at 37℃ overnight.

** The TE step washes off the chemical that is places on the filter to facilitate the lysis of bacterial clones and allows the DNA to stick. Failure to wash will inhibit transformation, but washing too long will cause DNA to elute and be lost.