Hieff NGS™ OnePot Pro DNA Fragmentation Module provides a new-generation enzymatic fragmentation reagent designed for Illumina® and MGI® high-throughput sequencing platforms . This product simplifies the operation process compared with the complex mechanical interruption and also greatly reduces the time and cost by performing fragmentation, end repair and dA-tailing of dsDNA in one reaction. The fragmentation products can immediately for adaptor ligation with no cleanup steps by using Hieff NGS™ Fast-Pace DNA Ligation Module (Cat#12607) or Hieff NGS™ Novel DNA Ligation Module (Cat#12626). Hieff NGS™ OnePot Pro DNA Fragmentation Module can work with a broad range of DNA inputs (animal, plant and microbial genomes), ranging from 500 pg-1μg in library preparation. Package Information No. Name 12619ES24 12619ES96 12619-A Smearase™ Buffer 240 μL 960 μL 12619-B Smearase™ Enzyme 120 μL 480 μL Note: 1. 12619 is a part of Hieff NGS® OnePot Pro DNA Library Prep Kit (Cat:12205). 2. The Smearase™ Enzyme is mixed with enzymes involved in end repair and dA-tailing. Cautions 1. This product is compatible with a range of 500 pg - 1 μg input DNA. High quality input DNA (A260/A280 = 1.8-2.0) is highly recommended. 2. High concentration of metal ion chelator or other salt remained in Input DNA may affect subsequent experiments, it is recommended to dilute DNA in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0). Table 1. Guideline for choosing the fragmentation time of conventional genomic DNA Main peak size of the insert fragment Fragmentation time Optimization range 600 bp 8 min 6-12 min 350 bp 10 min 8-14 min 250 bp 12 min 10-15 min 200 bp 15 min 13-18 min 150 bp 20 min 15-25 min Table 2. Guideline for choosing the fragmentation time of FFPE DNA Main peak size of the insert fragment Fragmentation time DIN* 250 bp 9-13 min > 8.0 250 bp 8-11 min 6.5-8.0 250 bp 4-8 min 4.2-6.5 250 bp 3-6 min 2.5-4.2 250 bp 9-13 min > 8.0 Note: *DIN is DNA Integrity Number, which a method to define the degree of FFPE DNA degradation by using Agilent 2200. 3. For conventional high-quality genomic DNA, fragmentation time refers to Table 1. This product is compatible with various samples with different GC content and has minimal bias. For the FFPE samples with different degrees of degradation, the fragmentation time refers to Table 2. Customers need to fine-tune the recommended fragmentation time in their own experimental system to achieve the best results. 4. The DNA can be digested to the required size according to the recommended fragmentation time. In order to ensure the high-quality and stable fragmentation effect, the fragmentation process should be operated on ice. 5. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.
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