Expand Long Template PCR System

for amplification of 5–20 kb targets from eukaryotic genomes Cat. No. 1 681 834    100 units

Cat. No. 1 681 842    2 x 250 units
Cat. No. 1 759 060    10 x 250 units

Description

Unique enzyme blend of Taq and Pwo DNA Polymerase. The system also includes PCR buffers for optimal amplification of different DNA fragment lengths:

PCR buffer 1 (10x conc.), with 17.5 mM MgCl₂ (for genomic targets up to 12 kb)
PCR buffer 2 (10x conc.), with 22.5 mM MgCl₂ (for 12–15 kb genomic targets)
PCR buffer 3 (10x conc.), with 22.5 mM MgCl₂, DMSO, and detergent (for 15–20 kb genomic targets and for targets amplified from difficult templates)

Application

Produces a high yield of long PCR products from episomal and genomic DNA.
The Expand Long Template PCR System is ideal for amplifying 5–20 kb fragments from human genomic DNA (or up to 40 kb fragments from DNA). The enzyme blend may be used for:

Mapping and sequencing whole genomes
Mapping intron/exon structures by amplifying cDNAs
Analyzing homologous recombination events
Characterizing cloned sequences carried in lambda-phage
Amplifying full-length cDNAs or long fragments of cDNA (RACE)

Expand Reverse Transcriptase is a genetically engineered version of M-MuLV Reverse Transcriptase. Point mutation within the RNase H sequence reduces the RNase H activity below detectable levels. This product is not for sale in the United States.

Application profile

Operating parameters

Starting sample: up to 500 ng genomic DNA or 10 ng lambda-DNA
pH optimum: 9.2
Divalent ion requirement: Mg²⁺
Temperature optimum (for elongation): 68°C

Experimental result

Amplification of a 29.7 kb and a 9.0 kb factor VIII region from genomic DNA with the Expand Long Template PCR System.

Key advantages

Increases fidelity of PCR amplification, because the Expand Long Template PCR System enzyme blend combines the proofreading activity of Pwo DNA Polymerase with the high processivity of Taq DNA Polymerase
Increases yield of PCR amplification, because the accuracy of the enzyme blend reduces the number of truncated amplification products formed
Improves the performance of long PCR, because the buffers included in the Expand Long Template PCR System are optimized for generation of certain length products

Roche Contact Information

Roche

Roche Diagnostics Corporation
P.O. Box 50414
9115 Hague Road
Indianapolis, IN 46250-0414 USA

Customer Service: 800-262-1640

Tech Support: 800-262-4911

Fax Number: 800-428-2883

Web Site: http://www.roche-applied-science.com

Expand Long Template PCR System