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Mesophilic DNA Polymerases: DNA Polymerase I; Klenow Fragment; Klenow Fragment, exo-; phi29 DNA Polymerase; T4 DNA Polymerase; T7 DNA Polymerase; Terminal Deoxynucleotidyl Transferase (TdT)

Mesophilic DNA Polymerases
Applications	Template	3'=>5' exo	5'=>3' exo	Strand displacement	Inactivation	Polymerase
Rolling circle amplification Multiple displacement amplification Whole genome amplification DNA sequencing Protein-primed DNA amplification In situ genotyping Recombination based-cloning Cell-free cloning of lethal DNA	DNA	++++	–	++++	Heating at 65°C, 10 min	phi29 DNA Polymerase
DNA labeling by nick-translation Second strand synthesis of cDNA	DNA	++	+	–	Heating at 75°C, 10 min	DNA Polymerase I
DNA blunting by fill-in 5'- overhangs Random-primed DNA labeling Labeling by fill-in 5'-overhangs of dsDNA DNA sequencing Site-specific mutagenesis of DNA with synthetic oligonucleotides Second strand synthesis of cDNA	DNA	++	–	++	Heating at 75°C, 10 min	Klenow Fragment
Random-primed DNA labeling Labeling by fill-in 5'-overhangs of dsDNA Strand displacement amplification DNA sequencing	DNA	–	–	++	Heating at 75°C, 10 min	Klenow Fragment, exo-
Blunting of DNA with 5'- or 3'-overhangs Blunting of PCR products with 3'-dA overhangs DNA labeling Site-directed mutagenesis Ligation-independent cloning of PCR products	DNA	++++	–	–	Heating at 75°C, 10 min	T4 DNA Polymerase
Purification of covalently closed circular DNA by removal of residual genomic DNA Primer extension reactions on long templates DNA 3'-end labeling Strand extensions in site-directed mutagenesis Fill-in blunting of 5'-overhang DNA Second strand synthesis of cDNA In situ detection of DNA fragmentation associated with apoptosis	DNA	++++	–	–	Heating at 75°C, 10 min	T7 DNA Polymerase
Production of synthetic homo-, heteropolymers Homopolymeric tailing of linear duplex DNA with any type of 3'-OH terminus Oligodeoxyribonucleotide and DNA labeling 5'-RACE In situ localization of apoptosis	ssDNA dsDNA ssRNA dsRNA	–	–	na	Heating at 70°C, 10 min	Terminal Deoxynucleotidyl Transferase (TdT)

- – not active
+ – active
na – not applicable
¹ – strand displacement activity of Pfu DNA Polymerase is temperature-dependent. It will perform strand displacement at temperatures higher than 60°C.