The SequalPrep™ Long PCR Kit with dNTPs provides the highest amplification success rate of any long range PCR kit tested across multiple organisms and platforms for non-optimized amplicons up to 20 kB with little to no optimization. Successful long range PCR with minimal optimization enables efficient PCR enrichment for a variety of applications including medical re-sequencing on next generation sequencing (NGS) platforms. PCR amplification is the traditional method to enrich target regions of the genome for genotyping and re-sequencing applications. To take advantage of the enhanced NGS processing capabilities, higher throughput enrichment methods have become necessary. Unfortunately, the potential efficiency gains from long range PCR using available commercial solutions have been offset with lower amplification success rates limiting enrichment to ∼5 kB regions. The SequalPrep™ Long PCR Kit with dNTPs overcomes these issues and provides the highest amplification success rate of any commercially available kit tested across multiple organisms and platforms for amplicons up to 20 kB with minimal optimization.
This kit can be used upstream of any NGS platform or for other genotyping applications. When combined with the SequalPrep™ Normalization Plate the SequalPrep™ (Sequence Equalization Sample Preparation) System is the first complete PCR enrichment and amplicon equalization solution that provides a viable and efficient enrichment method to meet NGS re-sequencing throughput capabilities. This NGS Sample Prep system significantly reduces PCR amplification and normalization complexity to allow for enrichment of regions up to 1MB in a 96 well plate format. Amplify and equalize with the ease of SequalPrep™ for consistently high coverage of your regions of interest.
研究用途にのみご使用ください。診断目的には使用できません。
仕様
エキソヌクレアーゼ活性3' - 5', 5' - 3'
フィデリティ(vs. Taq)4 X
ホットスタート内蔵ホットスタート
オーバーハングMixed
ポリメラーゼSequalPrep Long Polymerase
製品ラインSequalPrep
数量1,000 units
反応形態別のコンポーネント
出荷条件ドライアイス
サイズ(最終製品)20 kb or less
GC-Rich PCR Performance培地
反応速度スタンダード
Unit SizeEach
組成および保存条件
• 200 μL SequalPrep™ Long Polymerase (5 U/μL) • 1.2 ml SequalPrep™ 10X Reaction Buffer • 1.2 mL SequalPrep™ 10X Enhancer A • 1.2 mL SequalPrep™ 10X Enhancer B • 750 μL DMSO
Store at -20°C. DMSO may be stored at 4°C or room temperature.
よくあるご質問(FAQ)
What is the fidelity of the SequalPrep Long Polymerase in SequalPrep Long PCR Kit with dNTPs?
The SequalPrep Long PCR Kit with dNTPs demonstrates fidelity 3-4 times greater than recombinant Taq polymerase, similar to Invitrogen’s Platinum Taq High Fidelity Enzyme (by rpsl assay). The SequalPrep Long PCR Polymerase was compared to rTaq, Platinum Taq HiFi, & Roche's Expand Long Template PCR System. Relative fidelity:
• rTaq = 1
• Platinum Taq HiFi = 3.3
• Roche = 3.6
• SequalPrep Long with Enhancer A = 3.6
• SequalPrep Long with Enhancer B = 4.4
SequalPrep Long PCR Kit with dNTPs comes with several enhancers. What are they and when should I use them?
The two Enhancers were selected to help provide maximum success rates. We recommend setting up the initial reactions with 0.5X Enhancer A to provide maximum success rate. For any unsuccessful amplicons we recommend testing the following conditions to optimize amplification:
• 1.0X Enhancer A
• 0.5X Enhancer B
• 1.0X Enhancer B
What amount of input material do you recommend for SequalPrep Long PCR Kit with dNTPs?
We recommend 5-200 ng of human genomic DNA and 20-80 ng of mouse embryonic stem cells DNA. 50 ng of DNA template at a concentration of 50 ng/µl should be sufficient for most applications, although input amounts as low as 10 ng can be possible depending on the sample type, due to the high yields generated by the kit.