ExactRun High Fidelity proofreading DNA polymerase



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The ExactRun High-Fidelity DNA Polymerase offers robust performance and can be used for all PCR... more
Product information "ExactRun High Fidelity proofreading DNA polymerase"

The ExactRun High-Fidelity DNA Polymerase offers robust performance and can be used for all PCR applications including amplification of long fragments. Its unique features are a very high processivity, increased fidelity and speed (>6000pb per minute).

ExactRun DNA Polymerase is an ideal choice for cloning and can be used for long or difficult amplicons (20 kb for plasmid DNA or 7 for genomic DNA) with an error rate of about 50-fold lower than that of Taq DNA Polymerase or about 6-fold lower than that of Pfu DNA Polymerase.

Features:
- High Fidelity proofreading DNA polymearse.
- Extrem high amplification rate (>6000bp per minute).
- Error rate about 6-times less than that of Pfu or Pwo, about 50-times less compared to Taq.
- For long PCR fragments (up to 20 kb for plasmid DNA or up to 7 kb for genomic DNA.
- 5'-3' polymerase acitivity and 3'-5' exonuclease activity.
- generates blund ends.
- ExactRun DNA Polymerase is supplied with buffer and separate MgCl2.

More High-Fidelity Proofreading Polymerases from Genaxxon bioscience:

- M3002 Pwo Proofreading Polymerase >
- M3003 ReproFast Proofreading Polymerase >
- M3004 Pfu Proofreading Polymerase >
- M3012 ReproHot (KOD) Proofreading Polymerase >
- M3030 ExactRun (Phusion like) Proofreading Polymerase >

With our high quality dNTPs as Set (M3015.4100 and M3015.0250) > or Mix (M3016.1010) > or our DNA Ladders > and our favourable standard agarose (M3044) > we can offer additional products for your PCR.

More products around "ExactRun High Fidelity proofreading DNA polymerase"
Our comment on "ExactRun High Fidelity proofreading DNA polymerase"
- ExactRun is one of the most accurate thermostable polymerase available. - ExactRun DNA Polymerase possesses 5'-3' polymerase activity, 3'-5' exonuclease activity - ExactRun will generate blunt-ended products. - ExactRun DNA Polymerase is supplied with buffer and separate MgCl2.
Specifications: 2 units/µL 3´-5´ Exonuklease activity (proof-reading activity) more
 

Technical Data:

Specifications:
2 units/µL
3´-5´ Exonuklease activity (proof-reading activity)

application:

Proof-reading DNA polymerase for high fidelity and fast PCR.

Unit Definition:

One unit is defined as the amount of enzyme which will convert 10 nmoles of dNTPs to an acid-insoluble form in 30 min at 72°C under the assay conditions (25mM TAPS (tris-(hydroxymethyl)-methyl-amino-propanesulfonic acid, sodium salt) pH 9.3 (25°C); 50mM KCl; 2 mM MgCl2; 1mM b-mercaptoethanol; and activated calf thymus DNA as substrate.

Source

E.coli

Sicherheits Hinweise / Safety

Klassifizierungen / Classification

eclass-Nr: 34-16-04-90
Dokumente - Protokolle - Downloads more

Dokumente - Protokolle - Downloads

Here you will find information and further literature on ExactRun High Fidelity proofreading DNA polymerase. For further documents (certificates with additional lot numbers, safety data sheets in other languages, further product information) please contact Genaxxon biosience at: info@genaxxon.com or phone: +49 731 3608 123.

 
 
 
Dokumente - Protokolle - Downloads more

Dokumente - Protokolle - Downloads

Reaction conditions may differ from PCR protocols already in use, as ExactRun DNA Polymerase tends to need higher denaturation and annealing temperatures. Especially annealing temperature may differ significantly from that of Taq DNA polymerase.


- The amount of enzyme depends on the amount of template and the length of the amplicon. Usually 1 unit ExactRun per 50µL reaction volume is enough for good results, but the optimal amount can range from 0.25 to 2 units per 50µL reaction.
It is highly recommended not to exceed 2 units/50µL!

- When cloning fragments amplified with ExactRun DNA Polymerase, blunt end cloning is possible. If TA cloning is required please refer to page 5 (Using PCR-product for T/A-cloning).

- The concentration of Mg2+ is critical for PCR. Excessive Mg2+ stabilises the DNA double strand. Excess Mg2+ can also stabilize spurious annealing of primers to incorrect template sites and decrease specificity. Inadequate Mg2+ may lead to lower product yield. The optimal Mg2+ concentration depends on the dNTP concentration, the specific template DNA and the sample buffer. In general, the optimal Mg2+ concentration is ca. 1mM over the total dNTP concentration for standard PCR. If any component of the PCR reaction contains EDTA or EGTA, the apparent Mg2+ optimum may be shifted to higher concentrations.

- PCR additives / DMSO: Some publications recommend to add 3% DMSO for GC-rich templates, which shall aid to denature DNA. In some cases DMSO may also be required for supercoiled plasmids. Other additives as formamide, glycerol, and betaine can also be used together with ExactRun.
At high concentrations of DMSO the annealing temperature has to be reduced between 3°C and 6°C.

- Primer annealing: For primers >20nt use an annealing temperature of 5°C above the lower primer Tm.
For primers <20nt use an annealing temperature of 7°C above the lower primer Tm.
Anneal for 10 – 30 sec.
A 2-step PCR protocol is recommended when primer Tm values are at least 69°C.
Extension depends on template length and complexity We recommend an extension rate of 15 sec./kb for low complex DNA (plasmids, lambda DNA or BAC-DNA) and an extension rate of 30 sec/kb for high complex DNA (genomic). Sometimes extension rates have to be decreased down to 40 sec/kb.

    The amount of enzyme depends on the amount of template and the length of the amplicon. Usually 1 unit ExactRun per 50µL reaction volume is enough for good results, but the optimal amount can range from 0.25 to 2 units per 50µL reaction.

It is highly recommended not to exceed 2 units/50µL!

2.     When cloning fragments amplified with ExactRun DNA Polymerase, blunt end cloning is possible. If TA cloning is required please refer to page 5 (Using PCR-product for T/A-cloning).

3.     The concentration of Mg2+ is critical for PCR. Excessive Mg2+ stabilises the DNA double strand. Excess Mg2+ can also stabilize spurious annealing of primers to incorrect template sites and decrease specificity. Inadequate Mg2+ may lead to lower product yield. The optimal Mg2+ concentration depends on the dNTP concentration, the specific template DNA and the sample buffer. In general, the optimal Mg2+ concentration is ca. 1mM over the total dNTP concentration for standard PCR. If any component of the PCR reaction contains EDTA or EGTA, the apparent Mg2+ optimum may be shifted to higher concentrations.

4.     PCR additives / DMSO: Some publications recommend to add 3% DMSO for GC-rich templates, which shall aid to denature DNA. In some cases DMSO may also be required for supercoiled plasmids. Other additives as formamide, glycerol, and betaine can also be used together with ExactRun.
At high concentrations of DMSO the annealing temperature has to be reduced between 3°C and 6°C.

5.     Primer annealing: For primers >20nt use an annealing temperature of 5°C above the lower primer Tm.
For primers <20nt use an annealing temperature of 7°C above the lower primer Tm.
Anneal for 10 – 30 sec..
A 2-step PCR protocol is recommended when primer Tm values are at least 69°C.

Extension depends on template length and complexity We recommend an extension rate of 15 sec./kb for low complex DNA (plasmids, lambda DNA or BAC-DNA) and an extension rate of 30 sec/kb for high complex DNA (genomic). Sometimes extension rates have to be decreased down to 40 sec/kb.

 
 
 
Literatur .... more

Referenzen..

Hier finden Sie Artikel und Literaturzitate, in denen die Autoren auf die hohe Qualität dieses Genaxxonprodukts vertrauen.
Listed below are articles and references, in which the authors trust in the high quality of this Genaxxon product.

Quelle/Source: NCBI PubMed >

Structural Basis of the Interaction of MbtH-like Proteins, Putative Regulators of Nonribosomal Peptide Biosynthesis, with Adenylating Enzymes

Dominik A. Herbst, Björn Boll, Georg Zocher, Thilo Stehle, Lutz Heide

J Biol Chem. 2013 Jan 18; 288(3): 1991–2003. Published online 2012 Nov 28. doi: 10.1074/jbc.M112.420182

PMCID: PMC3548506

 
 
 
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