User Manual
Contents
1. lt gt MCLAB Molecular Cloning Laboratories User Manual Version 1 0 Revision Date 08 30 2013 Product name DNA polymerase thermotoga neapolitana Cat DPTN 100 DPTN 200 DPTN 300 DPTN OEM Description Due to its thermo stable nature DNA polymerase thermotoga neapolitana was identified 1 as an enzyme able to withstand the protein denaturing conditions high temperature required during PCR 2 Similar to Es cherichia coli DNA polymerase I but unlike Taq DNA polymerase Tne DNA polymerase contains both 3 gt 5 and 5 gt 3 exonulease activity Therefore it replaced the DNA polymerase from E coli originally used in PCR 3 Tag s optimum temperature for activity is 75 80 C with a half life of 9 minutes at 97 5 C and can replicate a 1000 bp strand of DNA in less than 10 seconds at 72 C 4 The DNA products have an A adenine overhangs at their 3 ends This may be useful in TA cloning whereby a cloning vector such as a plasmid that has a T thymine 3 overhang is used which complements with the A overhang of the PCR product thus enabling ligation of the PCR product into the plasmid vector Application PCR ordinary and high throughput Primer Extension Microarray Analysis Denaturing high performance liquid chromatography DHPLC Source Thermotoga neapolitana Tne DNA polymerase belongs to the DNA polymerase Pol family Recommended Reaction Conditions 94 C 1 minute2. gt 94 C 10 seconds gt 55 C 30 seconds gt 72 C 30 seconds for 25 cycles Recommended Storage Conditions 20 C Reference 1 Chien A Edgar DB Trela JM 1976 Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus J Bact 127 3 15507 PMC 232952 PMID 8432 2 Saiki RK et al 1988 Primer directed enzymatic amplification of DNA with a thermostable DNA poly merase Science 239 4839 48791 doi 10 1126 science 2448875 PMID 2448875 3 Saiki RK et al 1985 Enzymatic amplification of beta globin genomic sequences and restriction site anal ysis for diagnosis of sickle cell anemia Science 230 4732 1350 doi 10 1126 science 2999980 PMID 2999980 4 Lawyer FC et al 1993 High level expression purification and enzymatic characterization of full length Thermus aquaticus DNA polymerase PCR Methods Appl 2 4 27587 PMID 8324500 1 650 872 0245 www mclab com 1
Download Pdf Manuals
Related Search