Taq DNA polymerase (Robust buffer)

Product#: 02-02
$40.00
Size
Availability:
Ships in 24 hours

Taq DNA polymerase (Robust buffer)


Description

Thermus aquaticus DNA polymerase (Taq DNA polymerase) gene was expressed in E.coli in large quantities and highly purified. The enzyme has thermostable DNA polymerase activity and the MW is 94 kDa, same as that of the natural enzyme. 

This enzyme is suitable for PCR reactions; capable of amplifying DNA with various primers. 

Application

  1. High-throughput PCR
  2. Colony PCR
  3. Incorporation of dUTP, dITP, and fluorescence-labeled nucleotides
  4. Primer extension
  5. Addition of a single nucleotide (adenosine) at the 3’-blunt ends for cloning into TA vector.

Products
 
Product ID dNTP Size
02-02_02-012
- 200 units
02-02_02-012-5
- 5x200 units
02-02_02-002
+ 200 units
02-02_02-002-5
+ 5x200 units


General composition of PCR reaction mixture (total 50ul)

Taq DNA polymerase (5 units/ul) *0.25 ul
10 x Standard Buffer (Taq) 5 ul
2.5mM (each) dNTPs 4ul
Template <500ng
Primer 1 0.2~1.0uM (final conc.)
Primer 2 0.2~1.0uM (final conc.)
Sterile distilled water up to 50ul

*Use of excess amount of enzyme is not recommended. 


Storage Conditions

Taq DNA polymerase in 20mM Tris-HCl (pH 8.0), 100mM KCl, 0.1mM EDTA, 1mM DTT, 50% glycerol, 0.5% Tween20, 0.5% Igepal CA-630/
Store at -20℃

Concentration

5 units/ul, where one unit is defined as the amount of enzyme that can incorporate 10 nmols of total dNTPs into an acid-insoluble material in 30 minutes at 74℃ when activated salmon sperm DNA was used as template/primer.

Quality Assurance
Greater than 95% purity as determined by SDS-PAGE (CBB staining) (Fig.1) The absence of endonucleases and exonucleases was confirmed.

PCR Test 
Good amplification result was obtained in PCR reaction using λDNA as a template up to 14 kB (Fig.2). 

Reagents Supplied with Enzyme
1. 10 x Robust Buffer (Taq) 
02-012_LargPic.jpg 
Fig.1 SDS-PAGE analysis of Taq DNA polymerase 

Cautions for usage of Robust Buffer (Taq)
 
Robust Buffer induces maximum enzymatic activity. Therefore, cares should be taken to avoid production of undesirable smear bands in gel electrophoresis analysis by longer than optimal reaction time. We recommend about 5 to 10 seconds / kb elongation time for template up to 8 kb, and about 15 seconds / kb for up to 14 kb. We will recommend roughly the same elongation time to be set with 2-step PCR (shuttle PCR) and 3-step PCR. Extend the elongation time by short steps when amplification is not seen. The results of your experiments can be observed more rapidly by adopting 2-step PCR. 
 
Protocols for PCR
Examples of PCR coditions for the amplification of various sizes of λDNA (Results shown in Fig.2) 
02-012_fig1.jpg
Fig. 2 PCR products obtained by using Robust buffer (agarose gel electrophoresis) 
 
download.png

Satisfaction
Quality Rating
Value Rating
Style Rating
X