ELONGN-RO

Expand^™ Long Range dNTPack

suitable for PCR, hotstart: no, Long Range PCR

• Synonym(s): long template pcr, longe range pcr, nucleotide mixes and sets, pcr enzymes
• Enzyme Commission number: 2.7.7.7 (BRENDA, IUBMB)

Properties

• usage: sufficient for ≤1000 reactions (04829069001); sufficient for ≤200 reactions (04829042001); sufficient for >50 reactions (04829034001)
• Quality Level: 100
• feature: Long Range PCR; dNTPs included; hotstart: no
• packaging: pkg of 3,500 U (04829069001 [5x700 U]); pkg of 175 U (04829034001); pkg of 700 U (04829042001)
• manufacturer/tradename: Roche
• parameter: 68 °C optimum reaction temp.
• technique(s): PCR: suitable
• input: purified DNA
• storage temp.: −20°C

Description

General description

The Expand Long Range dNTPack is the second-generation version of the Expand Long Template PCR System, optimized and designed to consistently amplify PCR products from 5kb up to 25kb from genomic DNA (or up to 40kb from λ DNA templates). The supplied buffer can be used for all PCR product sizes. In addition, a buffer without MgCl₂, and separate vials of MgCl₂ and DMSO are included if optimization is needed. Expand Long Range dNTPack contains a special enzyme mix that consists of thermostable Taq DNA Polymerase and a thermostable DNA polymerase with proofreading activity. This powerful polymerase mixture produces a high yield of PCR product. Due to the inherent 3′–5′ exonuclease activity of the proofreading polymerase, Expand Long Range dNTPack copies DNA three times more accurately than Taq DNA Polymerase.

Specificity

Heat inactivation: > 92 °C

Application

The Expand Long Range dNTPack is optimized to efficiently amplify large genomic DNA fragments from 5kb to 25kb in combination with a threefold higher fidelity than Taq DNA Polymerase. The Expand Long Range dNTPack is ideally suited for:

• Genome mapping and sequencing
• Contig construction
• Characterization of cloned sequences in lambda phages or cosmids
• Cloning and analysis of eukaryotic genes
• Rapid identification and cloning of complete genes from genomic DNA using cDNA sequence as starting material
• Two-step RT-PCR

Use the specially designed buffer set, ultrapure PCR Nucleotide Mix, and optimized blend of thermostable DNA polymerases for:

• PCR
• Large-fragment amplification
• RT-PCR

For amplicon sizes of 20 to 35kb choose Expand 20kb PLUS PCR System, dNTPack.

Features and Benefits

Expand Long Range PCR System is ideal for genome mapping and sequencing, contig construction, characterization of cloned sequences in lambda phages or cosmids, and eukaryotic-gene cloning and analysis.
Amplify DNA fragments from 5 to 20kb from human genomic DNA, and 40kb from λDNA using supplied reaction buffers.
Use 3.75U for a standard 50μl PCR.
Each dNTPack contains 10mM additive-free sodium salt nucleotides as a ready-to-use mix.

• Amplify long templates

up to 25kb with high specificity, yield, and increased fidelity.

• Save time and resources.

Use one buffer, and fine-tune using the DMSO and MgCl₂ solution.

• Use this kit for all long template applications.

Obtain high yields of your desired fragment.

• Cost-effective.

Use the convenient premixed solution of PCR grade dNTPs.

Packaging

1 kit containing 6 components

Quality

Each lot is PCR function-tested using human genomic DNA and specific primers for a 9kb, 12kb, and 15kb fragment. The enzyme mix is tested for the absence of any contaminating activities, including endo- or exonucleases and nicking activity according to the current Quality Control procedures.

Unit Definition

Volume Activity: 5 U/μl

Preparation Note

Working concentration: Standard enzyme concentration: 3.5 U per 50 μl reaction

Other Notes

For life science research only. Not for use in diagnostic procedures.

Legal Information

Use of this product is covered by US patent claims and corresponding patent claims outside the US. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patent claims require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Expand is a trademark of Roche

Kit Components Only

• Expand Long Range Enzyme Mix
• Expand Long Range Buffer with 12.5 mM MgCl₂ 5x concentrated
• Expand Long Range Buffer without MgCl₂ 5x concentrated
• MgCl₂ Stock Solution 25 mM
• DMSO 100%
• PCR Nucleotide Mix

Safety Information

• Hazard Statements: H412
• Precautionary Statements: P273 - P501
• Hazard Classifications: Aquatic Chronic 3
• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 2
• Flash Point(F): 203.0 °F
• Flash Point(C): 95 °C