How do I “reconstitute” the cDNA after it arrives?
Store clones at 4°C until you are ready to use them. Clones are supplied as DNA (approximately 1.0 µg) spotted onto either a Whatman #1 filter disks, or dried within a 0.5ml screw-cap tube. Whatman Disk: To recover the DNA, put the filter paper into a 1.5 ml microcentrifuge tube. Add 100 µl of TE buffer (10 mM TRIS base, 1 mM EDTA, pH 8.0) to the microcentrifuge tube, vortex briefly, incubate at room temperature for 5 minutes, and repeat the vortex. Centrifuge the tube for a few seconds and then remove the filter paper from the tube. 0.5ml Tube: Centrifuge the tube BEFORE OPENING to ensure pellet is at the bottom of the tube. Add 50ul of ddH20 or TE buffer (as above) and vortex or flick the tube to ensure that the pellet disolves. For Both: Use 1-2 µl of supernatant for use in transfecting E. coli by electroporation or chemical means. Please do not try to use the DNA directly for any application other than to transform bacteria and prepare a large stock.
