Description

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 Fast-Pace DNA Ligation Module (Cat#12607) or Novel DNA Ligation Module (Cat#12626). 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:

12619 is a part of Hieff NGS® OnePot Pro DNA Library Prep Kit (Cat:12205).
The Smearase™ Enzyme is mixed with enzymes involved in end repair and dA-tailing.

Storage

OnePot Pro DNA Fragmentation Module is shipped with ice pack and can storage at -20℃ for 1 year.

Cautions

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.
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	15 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.

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.
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.
For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

Documents

Safety Data Sheet

12619-COMPONENT A-MSDS

12619-COMPONENT B-MSDS

Manuals

EN-12619 OnePot Pro DNA Fragmentation Reagent-HB220407.pdf

OnePot Pro DNA Fragmentation Module (end repair and dA-tailing plus)-12619ES