Optimize PCR amplification of long DNA amplicons up to 40 kb

• Fast, simple optimization starting with the complete kit
• Easily amplify DNA targets up to 40 kb long
• Reduce PCR-induced errors in these long amplicons with this high fidelity DNA polymerase mix

The MasterAmp™ Extra-Long PCR Kit is a complete system for successful and accurate one-step amplification of DNA sequences up to ~40 kb. The kit contains the MasterAmp™ Extra-Long DNA Polymerase Mix, as well as nine MasterAmp Extra-Long PCR 2X PreMixes with dNTPs, buffer, and varying amounts of MgCl₂ and the MasterAmp PCR Enhancer with Betaine.*

We recommend first using the complete kit (Cat. No. MHF9220) to test the different MasterMix Extra-Long 2X PreMixes (Step 1, Fig. 1) with each new template and primer set to identify the best performing one. One PreMix will amplify your long template and give consistent results in all subsequent amplifications (Step 2, Fig. 1). Once the optimal PreMix is identified continue to use that combination of MasterAmp Extra-Long 2X PreMix and DNA Polymerase for all subsequent amplifications using that template DNA and primer set.

Figure 1. Two steps to reliable and consistent PCR results.
STEP 1. Find the optimal MasterAmp™ Extra-Long PCR PreMix. Perform PCR with your template, primers, and the nine PreMixes.	STEP 2. Use the same MasterAmp™ Extra-Long PCR PreMix to obtain reliable, consistent PCR of the same sequence.
A. Amplification of a 20-kb region of lambda DNA using MasterAmp™ Extra-Long PCR 2X PreMixes (1-9). MasterAmp™ Extra-Long PCR PreMix 4 produced optimal results.	B. Subsequent PCR of a 20-kb lambda DNA region with the same primer pair and MasterAmp™ PreMix 4 gave consistent results, using the optimal PreMix determined in Step 1.

Figure 2. Extra-long amplification of lambda DNA using the MasterAmp™ Extra-Long PCR Kit. PCR conditions were optimized and MasterAmp Premix 4 was selected for PCR of 1 ng of lambda DNA in each sample. Results were analyzed on a 0.5% agarose gel. Amplicon sizes were 30 kb (lane 1), 35 kb (lane 2) and 40 kb (lane 3). Lane M, 5-Kb DNA ladder.

*Covered by issued and/or pending patents.

MasterAmp™ Extra-Long PCR Kit; Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser\"s own internal research. No other patent rights (such as 5´ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. 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.