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From 2011.igem.org

Agenda: Added Enzymes to E0840, J45119, and PSB1T3

(1)Vortex E0840, J45119, and PSB1T3.

(2)Add 1 microliter of SpeI enzyme and 1 microliter of PstI enzyme to 8 microliters of E0840 in order to cut both ends of the wanted DNA. Also add .5 microliters of BSA and 5 microliters.

(3)Add 1 microliter of EcoRI enzyme and 1 microliter of XbaI enzyme to J45119 in order to cut both ends of the wanted DNA. Also add .5 microliters of BSA and 5 microliters.

(4)Add 1 microliter of EcoRI enzyme and 1 microliter of PstI enzyme to PSB1T3 in order to cut both ends of the wanted DNA.

(5)Place E0840, J45119, and PSB1T3 in a water bath at 37 degrees C for 20 minutes so that all the parts are sufficiently cut.

(6)Assemble the new plasmid by combining 2 microliters of E0840, 2 microliters J45119, and 2 microliters PSB1T3 plasmids in one container. Also add 2 micro liters of buffer and 1 microliter of ligase.

(7)Place the new assembly of E0840, J45119, and PSB1T3 in the water bath at 37 degrees C for 20 minutes

(8)