Team:Mexico-UNAM-IPN/Horizontal-Transfer/29 September 2008


 * 1) Culture o/n of JM101 and JM109 strains.
 * 2) Dilution 1:100 in 30ml of sterile LB media. Incubate for 2.5-3hrs at 37°C.
 * 3) Thaw on ice for for 10 minutes. transfer to 12 vials.
 * 4) Centrifugate for 7min at 4°C at 3500rpm. Eliminate supernatant and decant. fill the vials with the culture and centrifugate again. Repeat ≈ 3 times.
 * 5) Resuspend the pellet in 1ml of sterile cold 10mM MgCl 2 in each vial. Add 500µl of 10mM MgCl 2 to eeach vial.
 * 6) Centrifugate at 3500 rpm for 7min at 40°C.
 * 7) Eliminate the supernatant and resuspend in 100µl of 50mM CaCl 2, Glycerol at 20%. Add 100µl of 50mM CaCl 2 , Glycerol at 20%. Incubate 1hr at 4°C.
 * 8) Transfer to 0.5ml eppendorf tubes and freeze.


 * JM101 : In blue the corresponding vials to this Escherichia colistrain.