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P2317

Sigma-Aldrich

10X PCR Buffer without MgCl2

Optimized for routine PCR without MgCl2

Synonym(s):

Magnesium-free PCR buffer

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About This Item

UNSPSC Code:
41106306
NACRES:
NA.52

form

liquid

technique(s)

PCR: suitable

color

colorless

application(s)

agriculture

shipped in

wet ice

storage temp.

−20°C

General description

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.

Features and Benefits

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

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

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Avoiding false positives with PCR.
Kwok, S., and Higuchi, R.
Nature, 229, 237-238 (1989)
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)
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)
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Protocols

dNTP Hot Start PCR Protocol

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

dNTP Hot Start PCR Protocol

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

dNTP Hot Start PCR Protocol

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

dNTP Hot Start PCR Protocol

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

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