merck millipore,默克密理博,RR013A,TaKaRa LA PCR Amplification Kit Version 2.1

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merck millipore,默克密理博,RR013A,TaKaRa LA PCR Amplification Kit Version 2.1
产品名称：TaKaRa LA PCR Amplification Kit Version 2.1
产品型号：RR013A

merck millipore,默克密理博,RR013A,TaKaRa LA PCR Amplification Kit Version 2.1

产品介绍

merck millipore,默克密理博,RR013A,TaKaRa LA PCR Amplification Kit Version 2.1

描述
产品目录编号	RR013A
品牌系列	Chemicon®
商名	Chemicon
描述	TaKaRa LA PCR Amplification Kit Version 2.1
概述	LA PCR Kit Ver.2.1. is designed and developed for Long and Accurate PCR Amplification of the target in any span length up to 48 kbp, employing a proprietary LA(long and accurate) PCR technology. Using TaKaRa LA Taq in conjunction with special reaction buffer (10x LA PCR Buffer II), it enables to generate longer and more accurate PCR products with greater yields. LA PCR Kit Ver.2.1 includes all the reagents required for optimal LA PCR in terms of efficient amplification, high fidelity and long PCR product yield. LA PCR technology will extend PCR applications, and is expected to be of particular value in facilitating genome analysis in various ways. Of course, this kit is also suitable to amplify short templates as well. Moreover, by using supplied specially designed buffer (2x GC Buffer I, II), this kit is applicable to amplify GC rich templates. Advantages: Longer and More Accurate PCR Amplification Extension to 20 kbp with Ease Up to 48 kbp Extension Capability Appliciable to GC Rich Template

产品信息
部件	*Amplifies 1,255 bp GC rich region of Control Template. TaKaRa LA Taq(5 units/μl) 25 μl dNTP Mixture (ea. 2.5 mM) 400 μl 10x LA PCR Buffer II (25 mM Mg2+ plus) 250 μl 10x LA PCR Buffer II (Mg2+ free) 250 μl MgCl2 (25 mM) 500 μl Control Template (100 ng/μl HT29 genomic DNA) 10 μl Control Primer LA3 (10 μM) 10 μl Control Primer LA4 (10 μM)* 10 μl --Hind III digest MW Marker (100 ng/μl) 20 μl 2x GC Buffer I (5 mM Mg2+ plus) 1.25 ml 2x GC Buffer II (5 mM Mg2+ plus) 1.25 ml Control Primer GC1 (10 μM) 10 μl Control Primer GC2 (10 μM) 10 μl *Amplifies 17.5 kbp region of Control Template.

产品信息

部件

**Amplifies 1,255 bp GC rich region of Control Template.
TaKaRa LA Taq(5 units/μl) 25 μl
dNTP Mixture (ea. 2.5 mM) 400 μl
10x LA PCR Buffer II (25 mM Mg2+ plus) 250 μl
10x LA PCR Buffer II (Mg2+ free) 250 μl
MgCl2 (25 mM) 500 μl
Control Template (100 ng/μl HT29 genomic DNA) 10 μl
Control Primer LA3 (10 μM)* 10 μl
Control Primer LA4 (10 μM)* 10 μl
--Hind III digest MW Marker (100 ng/μl) 20 μl
2x GC Buffer I (5 mM Mg2+ plus) 1.25 ml
2x GC Buffer II (5 mM Mg2+ plus) 1.25 ml
Control Primer GC1 (10 μM)** 10 μl
Control Primer GC2 (10 μM)** 10 μl
*Amplifies 17.5 kbp region of Control Template.

应用
主要应用	DNA Amplification
应用说明	DNA amplification by Polymerase Chain Reaction (PCR). PCR condition (an example) When amplifying 1 kbp DNA fragment 30 cycles of : 98°C 10sec, 55°C 30sec, 72°C 1min or 30 cycles of 98°C 10sec, 68°C 1min. Note: Denaturation conditions vary depending on the cycler and tube. It is recommended for 10-30sec at 94°C, or 1-10sec at 98°C. General reaction mixture for PCR (total 100μL) TaKaRa Ex Taq™ 0.5μL 10X Ex Taq™ Buffer 10μL dNTP Mixture (2.5mM each) 8μL Template <1μg Primer 1 20-100pmol (final conc. 0.2-1.0μM) Primer 2 20-100pmol (final conc. 0.2-1.0μM) Sterilized distilled water up to 100μL PCR products: As most PCR products amplified with TaKaRa ex Taq™ have one A added at the 3'-termini, the obtained PCR product can be directly used for cloning into a T-vector. Also, it is possible to clone the product in blunt-end vectors after blunting and phosphorylation of the end. Cool Start Method: Minimizes the amplification of non-specific bands in pCR and achieves more accurate amplification. This is a simpler method without the need for special enzymes nor additional reagents. Protocol: 1. Keep all reagents on ice until use. 2. Prepare the reaction mixture on ice. The adding order of reagents does not influence results. The result will not be affected even when the micutre is left on ice 30 min before thermal cycling. 3. Set a thermal cycler ready to start with the designated program. There is no need to change PCR conditions especially for Cool Start. 4. Set the tubes in thermal cycler and start cycling immediately. Japan patent 2576741 for Cool Start Method is owned by Shimadzu Corporation.

应用

主要应用

DNA Amplification

应用说明

DNA amplification by Polymerase Chain Reaction (PCR).

PCR condition (an example)

When amplifying 1 kbp DNA fragment

30 cycles of : 98°C 10sec, 55°C 30sec, 72°C 1min or

30 cycles of 98°C 10sec, 68°C 1min. Note: Denaturation conditions vary depending on the cycler and tube. It is recommended for 10-30sec at 94°C, or 1-10sec at 98°C.

General reaction mixture for PCR (total 100μL)

TaKaRa Ex Taq™ 0.5μL

10X Ex Taq™ Buffer 10μL

dNTP Mixture (2.5mM each) 8μL

Template <1μg

Primer 1 20-100pmol (final conc. 0.2-1.0μM)

Primer 2 20-100pmol (final conc. 0.2-1.0μM)

Sterilized distilled water up to 100μL

PCR products: As most PCR products amplified with TaKaRa ex Taq™ have one A added at the 3'-termini, the obtained PCR product can be directly used for cloning into a T-vector. Also, it is possible to clone the product in blunt-end vectors after blunting and phosphorylation of the end.

Cool Start Method*: Minimizes the amplification of non-specific bands in pCR and achieves more accurate amplification. This is a simpler method without the need for special enzymes nor additional reagents.

Protocol:

1. Keep all reagents on ice until use.

2. Prepare the reaction mixture on ice. The adding order of reagents does not influence results. The result will not be affected even when the micutre is left on ice 30 min before thermal cycling.

3. Set a thermal cycler ready to start with the designated program. There is no need to change PCR conditions especially for Cool Start.

4. Set the tubes in thermal cycler and start cycling immediately.

* Japan patent 2576741 for Cool Start Method is owned by Shimadzu Corporation.

生物信息
浓缩	5 units/μL

产品使用声明
销售限制	This product is only available for sale in the United States.
使用声明	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.