11811002001

Expand^™ 20 kb^PLUS PCR System

sufficient for ≤40 reactions, pkg of 200 U, suitable for PCR

• Synonym(s): long range PCR | long template PCR
• Enzyme Commission number: 2.7.7.7 (BRENDA, IUBMB)
• UNSPSC Code: 41106300

Properties

• usage: sufficient for ≤40 reactions
• feature: Long Range PCR; dNTPs included: no; hotstart: no
• packaging: pkg of 200 U
• manufacturer/tradename: Roche
• parameter: 68 °C optimum reaction temp.
• technique(s): PCR: suitable
• input: purified DNA
• shipped in: dry ice
• storage temp.: −20°C

Description

General description

This PCR system is an improvement of the Barnes Technology and shows good performance for the amplification of fragments longer than 20 kb.
The Expand 20 kb^PLUS PCR System is composed of a special enzyme mix containing thermostable Taq DNA polymerase and Tgo DNA polymerase, a thermostable DNA polymerase with proofreading activity. This powerful polymerase mixture and associated buffer system is designed to give a high yield of PCR product when fragments longer than 20 kb need to be amplified.
T/A cloning can be used for fragments produced by this kit.
To obtain long PCR products, first evaluate length and purity of the DNA template. Handling all genomic DNA templates with care. Genomic DNA template should be larger than 50 kb, as determined with agarose gel electrophoresis.

The Expand 20 kb^PLUS PCR System contains human control DNA and human control ß-globin primers which allow the amplification of a 23 kb fragment.
These reagents may serve as a control reaction but can also be used to test the quality of human template DNA ´s and/or the respective primer pairs.

T/A cloning can be applied.
For a standard 50 μl PCR, we recommend using 5 U of the enzyme blend.

Contents:

• Expand 20 kb^PLUS Enzyme Mix
• Expand 20 kb^PLUS Reaction Buffer, 10x concentrated with 27.5 mM MgCl₂
• MgCl₂ Solution, 25 mM
• Human Genomic DNA
• Human β-globin Control Primer, forward (HβG forward)
• Human β-globin Control Primer, reverse (HβG reverse)

Application

Use the Expand^™ 20 kb^PLUS PCR System to amplify genomic DNA fragments up to 35 kb. The Expand 20 kb^PLUS PCR System features a specifically optimized buffer and enzyme-blend mixture to amplify extra-long pieces of DNA (over 20 kb). Control primers and DNA produce fragments of 23 kb in length.

• PCR
• RT-PCR
• Large-fragment amplification

Use the Expand 20 kb^PLUS PCR System, dNTPack with ready-to-use PCR Nucleotide Mix.

Features and Benefits

• Go the limit.

The buffer and enzyme-blend amplify products over 20 kb DNA.

• Easy PCR monitoring.

Human genomic DNA and human control ß-globin primers amplify a 23 kb fragment.

• Test template quality.

Control reagents also test template quality and primer pair performance.

Packaging

1 kit containing 6 components

Quality

Each lot of Expand 20 kb^PLUS PCR System is function-tested in PCR using human genomic DNA and specific primers for β-globin to amplify a 23 kb fragment.

Unit Definition

Volume Activity: 5 U/μl

Other Notes

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

Legal Information

Use of this product is covered by US Patent No. 6,127,155 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 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. Human and veterinary diagnostic uses under Roche patent claims require a separate license from Roche.  All uses other than internal research and human and veterinary diagnostic uses under Roche patent claims require a separate license from Thermo Fisher Scientific. Further information on purchasing licenses from Roche may be obtained by contacting the Licensing Department of Roche Molecular Systems, Inc., 4300 Hacienda Drive, Pleasanton, California 94588, USA or Roche Diagnostics GmbH, Sandhofer Strasse 116, 68305 Mannheim, Germany.  Further information on purchasing licenses from Thermo Fisher Scientific may be obtained by contacting the Licensing Department of  Thermo Fisher Scientific, 5791 Van Allen Way, Carlsbad, California 92008, USA.

Expand is a trademark of Roche

Kit Components Only

• Expand 20 kb^PLUS Enzyme Mix
• Expand 20 kb^PLUS Reaction Buffer, with 27.5 mM MgCl₂ 10x concentrated
• MgCl₂ Stock Solution 25 mM
• Human Genomic DNA
• Human β-globin Control Primer forward, (HβG forward)
• Human β-globin Control Primer reverse, (HβG reverse)

Safety Information

• hcodes: H412
• pcodes: P273 - P501
• Hazard Classifications: Aquatic Chronic 3
• Storage Class: 12 - Non Combustible Liquids
• wgk_germany: WGK 2
• flash_point_f: does not flash
• flash_point_c: does not flash