TOPO TA Cloning (Invitrogen)

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Kit Contents and StorageShipping andStorageTOPO TA Cloning ® Kits are shipped on dry ice. Each kit contains a box with TOPO TACloning ® reagents (Box 1) and a box with One Shot ® Chemically Competent orElectrocomp cells (Box 2). Store Box 1 at -20°C and Box 2 at -80°C.Types of TOPO TACloning ® KitsTOPO TA Cloning ® Kits are available with either pCR ® 2.1-TOPO ® or pCR ® II-TOPO ®and either DH5α -T1 R , TOP10 or TOP10F´ One Shot ® Chemically Competent cells orTOP10 One Shot ® Electrocomp cells (see page vi for the genotypes of the strains).Product Reactions One Shot ® Cells Type of Cells Catalog no.TOPO TA Cloning ® Kit 20 TOP10 chem. competent K4500-01(containing pCR ® 2.1-TOPO ® ) 40 TOP10 chem. competent K4500-4020 DH5α-T1 R chem. competent K4520-0140 DH5α-T1 R chem. competent K4520-4020 TOP10F´ chem. competent K4550-0140 TOP10F´ chem. competent K4550-4020 TOP10 electrocompetent K4560-01TOPO TA Cloning ® Kit Dual Promoter 20 TOP10 chem. competent K4600-01(containing pCR ® II-TOPO ® ) 40 TOP10 chem. competent K4600-4020 DH5α-T1 R chem. competent K4620-0140 DH5α-T1 R chem. competent K4620-4020 TOP10F´ chem. competent K4650-0140 TOP10F´ chem. competent K4650-4020 TOP10 electrocompetent K4660-01continued on next pageiv
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
Page 1: TOPO TA Cloning ®Version N07260125
Page 6 and 7: Kit Contents and Storage, continued
Page 8 and 9: Purchaser NotificationCloningTechno
Page 10 and 11: Overview, continuedExperimentalOutl
Page 12 and 13: TOPO ® Cloning Reaction and Transf
Page 18 and 19: Map of pCR ® 2.1-TOPO ®pCR ® 2.1
Page 20 and 21: TOPO TA Cloning ® Control Reaction
Page 22 and 23: TOPO TA Cloning ® Control Reaction
Page 24 and 25: Purifying PCR Products, continuedLo
Page 26 and 27: RecipesLB (Luria-Bertani)Medium and
Page 28 and 29: Technical Service, continuedEmergen

TOPO TA Cloning (Invitrogen)

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