Multiplex PCR 5X Master Mix |NEB酶试剂 New England Biolabs

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Multiplex PCR 5X Master Mix

Multiplex with flexibility and minimal optimization

  • Simultaneously detect two or more products in a single reaction
  • Enhanced buffer formulation for optimal performance 
  • 5X master mix format for easy reaction setup – just add template DNA and primers

Multiplex PCR 5X Master Mix |

库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
M0284S 5 X 100 reactions (50 µl) ¥2,729.00 有 无 无 无 无
如何订购
Please enter a quantity for at least one size

产品信息

Multiplex PCR can simultaneously detect two or more products in a single reaction. There is an increasing demand for multiplex PCR techniques in assays conducted in research laboratories and forensic/diagnostic genotyping assays (1,2). Multiplex PCR can also be used for semi-quantitative gene expression analysis using cDNA templates. The NEB Multiplex PCR 5X Master Mix is an easy-to-use solution featuring high quality recombinant Taq DNA Polymerase. The mix is optimized for high yield and robust performance. Its performance is illustrated in a 15-plex PCR using human genomic DNA (Figure 1) and an 8-plex PCR using cDNA products as templates (Figure 2). The 5X formulation allows the user maximal input of primers, template DNAs and additional components.

Figure 1
Multiplex PCR 5X Master Mix |
15-plex PCR using varying amounts of human genomic DNA. 1X Multiplex PCR 5X Master Mix was used with 0.15 μM of each primer. The cycling conditions were 95°C for 1 minute, 35 cycles of 95°C for 20 seconds, 60°C for 1 minute and 68°C for 2 minutes. Marker M is the 1 kb Plus DNA Ladder (NEB #N3200).
Figure 2
Multiplex PCR 5X Master Mix |
8-plex PCR using cDNA products from 1 ng human spleen total RNA. Cycling conditions were 95°C for 1 minute, 30 cycles of 95°C for 20 seconds, 60°C for 30 seconds and 68°C for 2 minutes. Amplicon lengths are indicated next to gel.
Figure 3
Multiplex PCR 5X Master Mix |
Comparison of PCR product yields between single-plex and multiplex PCR reactions. Figure A shows amplification of human genomic DNA fragments (sizes are listed to the left of the gel). Lane 1 is a 3-plex reaction, Lanes 2-4 are single-plex reactions, and Lanes 5–6 are 2-plex reactions; Lane 7 is the 1 kb DNA Ladder (NEB #N3232 ). Figure B shows the results of expression analysis of five mRNAs using cDNA templates (first strand synthesis was carried out using the ProtoScript Kit with human spleen total RNA); Lane 1 is a 5-plex PCR from cDNA reactions, while Lanes 2–6 are single-plex PCR reactions; Lane 7 is the 100 bp DNA Ladder (NEB #N3231).

产品来源

An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1.

产品类别:
Taq DNA Polymerase Products,
Master Mixes Products

应用:
Multiplex PCR,
Specialty PCR,
PCR

  • 产品组分信息

    产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • M0284S     -20    
        Multiplex PCR 5X Master Mix M0284SVIAL -20 1 x 1 ml 5 X

  • 特性和用法

    1X 预混液组分

    20 mM Tris-HCl
    50 mM KCl
    30 mM NH4Cl
    2.5 mM MgCl2
    0.3 mM dNTPs
    3.2% Glycerol
    0.08% IGEPAL® CA-630
    0.07% Tween® 20
    67 units/ml Taq DNA Polymerase
    pH 8.9 @ 25°C

    热失活

    否

    分子量

    理论上的: 94000 daltons

    5′ – 3′ 核酸外切酶

    Yes

    3′ – 5′ 核酸外切酶

    No

    链置换

    No

    产物末端

    • Single-base 3´ Overhangs

    单位活性检测条件

    1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA.

    Unit Definition:
    One unit of Taq DNA Polymerase is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C.

  • 优势和特性

    Features

    • Multiplex with Taq

  • 注意事项
    1. Use high quality primers (desalted or HPLC purified).
    2. Accurately quantify and adjust primer concentrations to 50 μM in 0.5X TE Buffer.
    3. Individually test the PCR primer pairs, preferentially in a temperature-gradient PCR machine.
    4. Mix all primers equally at 1 μM in 0.5X TE buffer.
    5. Keep at -20°C for long term storage.
    6. Multiplex PCR 5X Master Mix is stable for 15 freeze-thaw cycles when stored at -20°C.
    7. Multiplex PCR 5X Master Mix is stable at 4°C for three months. So, for frequent use, an aliquot may be kept at 4°C.

  • 参考文献
    1. Beggs et al. (1990). Hum. Genet.. 86, 45-48.
    2. Krenke et al. (2002). J. Forensic Sci.. 47, 773-785.
    3. Sun, Y. et al. (1993). Biotechniques. 15, 372-374.

操作说明、说明书 & 用法

  • 操作说明
    1. Multiplex PCR Guidelines for Multiplex PCR 5X Master Mix
    2. Quick Protocol for Multiplex PCR 5X Master Mix

  • 使用指南
    • Activity of Restriction Enzymes in PCR Buffers
    • Guidelines for PCR Optimization with Thermophilic DNA Polymerases

工具 & 资源

  • Web 工具
    • Tm Calculator

FAQs & 问题解决指南

  • FAQs
    1. What is the stability of Multiplex PCR 5X Master Mix?
    2. How should I set up a PCR using Multiplex PCR 5X Master Mix?
    3. What type of DNA ends result from a primer extension reaction or a PCR using Taq DNA Polymerase?
    4. Does the presence of Ca2+ inhibit PCR?

  • 问题解决指南
    • PCR Troubleshooting Guide

  • 实验技巧
    Test each primer set individually before combining into a multiplex reaction. The concentration of some primer pairs may need to be adjusted to achieve consistent and robust amplification across all amplicons in the mix.

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

    Multiplex PCR 5X Master Mix | Powered by Bioz See more details on Bioz

质控、安全 & 法规

  • 质控分析
    每一新批次的 NEB 产品都要经过质控，以满足指定的质量标准，对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息，可从此处查阅。

  • 产品说明与变更通知

    产品说明与变更通知

    产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]

    • M0284S_v1
    • M0284S_v2

  • CoA
    CoA 文件包含单个批次产品的储存温度、有效期和质控，这些文件的命名规则如下： [货号]_[规格]_[版本]_[Lot#]

    • M0284S_v1_0201703
    • M0284S_v1_0221803
    • M0284S_v1_10018521
    • M0284S_v1_10026399
    • M0284S_v1_10035199
    • M0284S_v1_10049756
    • M0284S_v1_10056678
    • M0284S_v1_10059631
    • M0284S_v2_10087106
    • M0284S_v2_10092903
    • M0284S_v2_10122832
    • M0284S_v2_10146119

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • Multiplex PCR 5X Master Mix