TOPO TA Cloning (Invitrogen)

Kit Contents and Storage, continuedTOPO TA Cloning ®ReagentsTOPO TA Cloning ® reagents (Box 1) are listed below. Please note that the user mustsupply Taq polymerase. Store Box 1 at -20°C.Item Concentration AmountpCR ® 2.1-TOPO ® orpCR ® II-TOPO ®10 ng/µl plasmid DNA in:;50% glycerol50 mM Tris-HCl, pH 7.4 (at 25°C)1 mM EDTA1 mM DTT0.1% Triton X-100100 µg/ml BSAphenol red20 µl10X PCR BufferSalt SolutiondNTP Mix100 mM Tris-HCl, pH 8.3 (at 42°C)500 mM KCl25 mM MgCl 20.01% gelatin100 µl1.2 M NaCl0.06 M MgCl 250 µl12.5 mM dATP; 12.5 mM dCTP12.5 mM dGTP; 12.5 mM dTTPneutralized at pH 8.0 in water10 µlM13 Forward (-20) Primer 0.1 µg/µl in TE Buffer 20 µlM13 Reverse Primer 0.1 µg/µl in TE Buffer 20 µlControl Template 0.1 µg/µl in TE Buffer 10 µlControl PCR Primers 0.1 µg/µl each in TE Buffer 10 µlSterile Water -- 1 mlSequence ofPrimersThe table below describes the sequence and pmoles supplied of the sequencing primersincluded in this kit.Primer Sequence pMoles SuppliedM13 Forward (-20) 5´-GTAAAACGACGGCCAG-3´ 407M13 Reverse 5´-CAGGAAACAGCTATGAC-3´ 385continued on next pagev