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P2317

Sigma-Aldrich

10X PCR Buffer without MgCl2

Optimized for routine PCR without MgCl2

Synonyme(s) :

Magnesium-free PCR buffer

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About This Item

Code UNSPSC :
41106306
Nomenclature NACRES :
NA.52

Forme

liquid

Technique(s)

PCR: suitable

Couleur

colorless

Application(s)

agriculture

Conditions d'expédition

wet ice

Température de stockage

−20°C

Description générale

10X PCR Buffer II was tested at a final concentration of 1X (10mM Tris-HCl, pH 8.3 at 25 °C, 50mM KCl), in reactions containing 1-4mM MgCl2, each dNTP at 200 μM, primers defining an approximately 500 base pair region of λ DNA at 1.0μM each, λ DNA template at 1ng/100μL, and Taq DNA polymerase at 2.5 units/100μL. The reaction underwent 25 cycles of 94 °C to denature the double stranded DNA, 55 °C to anneal the DNA segments and 72 °C to extend the DNA segments. Following electrophoresis of the reaction products in 1.5% agarose gel, a single band of approximately 500 base pairs was visualized for PCRs containing 1-1.5mM MgCl2.

Application

10X PCR Buffer without MgCl2 has been used as a component of polymerase chain reaction (PCR) amplification mixture for the mycotoxigenic mould DNA, parasite DNA and Fusarium oxysporum DNA amplification.It has also been used as a component of the PCR mix for the amplification of nuclear ribosomal internal transcribed spacer (ITS2) and the partial ribulose-1,5 bisphosphate carboxylase/oxygenase large subunit rbcl gene for molecular and morphological characterization of Ulva sp from the Persian Gulf.
10× PCR Buffer without MgCl2 has been used with Sigma′s PCR enzymes.

Caractéristiques et avantages

  • This product is tested for the absence of DNase and RNase.
  • Suitable for use with magnesium chloride.

Code de la classe de stockage

12 - Non Combustible Liquids

Classe de danger pour l'eau (WGK)

WGK 1

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable


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Consulter la Bibliothèque de documents

A simple method of preparing plant samples for PCR.
H Wang et al.
Nucleic acids research, 21(17), 4153-4154 (1993-08-25)
Andrée-Anne Dussault et al.
Biological procedures online, 8, 1-10 (2006-02-01)
Real-time polymerase chain reaction (PCR) constitutes a significant improvement over traditional end-point PCR, as it allows the quantification of starting amounts of nucleic acid templates, in real-time. However, quantification requires validation through numerous internal controls and standard curves. We describe
Molecular characterization of Fusarium oxysporum f. sp. cubense isolates from banana
Das A, et al.
Pest Management Science, 18(2), 171-178 (2012)
Molecular and morphological characterisation of Ulva chaugulii, U. paschima and U. ohnoi (Ulvophyceae) from the Persian Gulf, Iran
Pirian K, et al.
Botanica Marina, 59(2-3), 147-158 (2016)
Avoiding false positives with PCR.
Kwok, S., and Higuchi, R.
Nature, 229, 237-238 (1989)

Protocoles

Hot Start dNTPs block DNA polymerase until heat activation, enhancing PCR specificity.

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