P2317
10X PCR Buffer without MgCl2
Optimized for routine PCR without MgCl2
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Synonym(s):
Magnesium-free PCR buffer
UNSPSC Code:
41106306
NACRES:
NA.52
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Quality Level
form
liquid
technique(s)
PCR: suitable
color
colorless
application(s)
agriculture
shipped in
wet ice
storage temp.
−20°C
Related Categories
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.
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
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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)
Mould incidence and mycotoxin contamination in freshly harvested maize kernels originated from India
Mudili V, et al.
Journal of the Science of Food and Agriculture, 94(13), 2674-2683 (2014)
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
A simple method of preparing plant samples for PCR.
H Wang et al.
Nucleic acids research, 21(17), 4153-4154 (1993-08-25)
Protocols
Hot Start dNTPs block DNA polymerase until heat activation, enhancing PCR specificity.
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