Recombinant Taq DNA Polymerase Proteinused for PCR various applications, CAS No.9012-90-2

Item Number

A156186

• Product Name: Recombinant Taq DNA Polymerase Proteinused for PCR various applications, CAS No.9012-90-2
• Synonyms: DNA polymerase I | thermostable | DNA-directed DNA polymerase
• Grade: EnzymoPure™
• Specifications & Purity: EnzymoPure™, ≥95%, Lot by Lot
• Bioactivity: Taq DNA polymerase is a heat-resistant enzyme that synthesizes DNA from a single-stranded template in the presence of dNTP and primers, with 5´→3´ DNA polymerase activity and 5´→3´ exonuclease activity.
• Expression System: E. coli
• Species: Thermus aquaticus
• Amino Acids: 1-832 aa
• Sequence: MRGMLPLFEPKGRVLLVDGHHLAYRTFHALKGLTTSRGEPVQAVYGFAKSLLKALKEDGDAVIVVFDAKAPSFRHEAYGGYKAGRAPTPEDFPRQLALIKELVDLLGLARLEVPGYEADDVLASLAKKAEKEGYEVRILTADKDLYQLLSDRIHVLHPEGYLITPAWLWEKYGLRPDQWADYRALTGDESDNLPGVKGIGEKTARKLLEEWGSLEALLKNLDRLKPAIREKILAHMDDLKLSWDLAKVRTDLPLEVDFAKRREPDRERLRAFLERLEFGSLLHEFGLLESPKALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKALRDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEEAGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALSLEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAAVLEALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSSSDPNLQNIPVRTPLGQRIRRAFIAEEGWLLVALDYSQIELRVLAHLSGDENLIRVFQEGRDIHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERYFQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPVQGTAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYPLAVPLEVEVGIGEDWLSAKE
• Protein Tag: N-His
• Protein Length: Protein fragments
• Source: Recombinant
• Predicted molecular weight: 96 kDa
• SDS-PAGE: 96 kDa

Product Description

Purity:

>95%, by SDS-PAGE visualized with Coomassie® Blue Staining.

Taq DNA Polymerase is a thermostable enzyme that synthesizes DNA from single-stranded templates in the presence of dNTPs and a primer. The enzyme consists of a single polypeptide with a molecular weight of 94 kDa. It has a 5´→3´ DNA polymerase activity and a 5´→3´ exonuclease activity.

Source：
Recombinant enzyme is purified from the cloned Thermus aquaticus DNA polymerase gene expressed in E. coli.

Unit definition

One unit of Taq DNA Polymerase is the amount of enzyme required to incorporate 10 nmoles of deoxyribonucleotide into DNA in 30 min at 74°C.

Product content：

1. Taq DNA Polymerase ( 5 U/μL )

2. 10×Taq Buffer（Mg2+Plus）：200mM Tris-HCl,500mM KCl,20mM MgCl2,1mM DTT,pH9.3

PCR Reaction Setup：

Component	50-μL rxn
5×Taq Buffer(Mg2+ Plus)	10μL
dNTP Mixture(2.5mM each)	4μL
Template DNA	＜500 ng
Primer F	0.2-1.0 μM
Primer R	0.2-1.0 μM
Taq DNA Polymerase(5U/μL)	0.2μL
ddH2O	Up to 50μL

Incubate reactions in a thermal cycler：

Step	Temperature（℃）	Time	Cycles
Initial Denaturation	95	5min	1
Denature	95	30s	25-35
Anneal	~55(depending on primer Tm)	30s
Extend	72	1min/kb
Final Extension	72	10min	1
Hold	4	indefinitely	1

Optimization Strategies：

Different PCR reaction conditions, including temperature, time and number of cycles, should be set according to different template, primer, length of PCR product and GC content.

The time setting of STEP4(extension) should be set according to the length of PCR products, usually the extension time of each kb product is 1min.

For initial PCR, the number of cycles can be set to 35 to ensure that the expected PCR product can be amplified as much as possible. The number of PCR reaction cycles that need to be semi-quantitative or quantitative must be properly optimized so that the PCR reaction does not reach a plateau.