Beschrijving
Hieff™ UltraShear Methylated DNA Fragmentase is an optimized enzyme mixture specifically designed for DNA fragmentation prior to library preparation. Unlike conventional fragmentation enzymes, this formulation preserves DNA methylation marks. Compared to traditional sonication, this product causes minimal damage to DNA templates, results in lower sample loss, and eliminates the cumbersome sonication step—making the fragmentation process simpler and more efficient. It is suitable for 5–250 ng of standard genomic DNA from animals, plants, or microorganisms, and is also compatible with FFPE-derived DNA.
Features
Methylation-Friendly: Preserves methylation marks and is fully compatible with methylation library preparation.
Gentle and Efficient: Minimizes template damage and sample loss while enabling one-step fragmentation.
Broad Compatibility: Works with 5–250 ng DNA from plants, animals, microbes, and FFPE samples; no sonication required and automation-friendly.
Components
|
Components No. |
Name |
12218ES08 (8 T) |
12218ES24 (24 T) |
12218ES96 (96 T) |
|
|
12218-A |
|
UltraShear Reaction Buffer |
56 μL |
168 μL |
672 μL |
|
12218-B |
|
UltraShear Enzyme |
48 μL |
144 μL |
576 μL |
|
12218-C |
|
UltraShear Reagent |
8 μL |
24 μL |
96 μL |
Storage
This product should be stored at -25~-15℃ for 1 year.
Application
dsDNA fragmentation; Methylation library preparation
Instructions
1. Thaw reagents listed in Table 1, invert to mix, and keep on ice.
2. Prepare the reaction mix for DNA fragmentation on ice as described in Table 1.
Table 1. DNA Fragmentation Reaction Setup
|
Component |
Volume (μL) |
|
Input DNA |
X |
|
UltraShear Reaction Buffer |
7 |
|
UltraShear Enzyme |
6 |
|
ddH₂O |
Up to 60 |
3. Gently mix by pipetting or light vortexing, then briefly centrifuge to collect liquid at the bottom of the tube.
4. Place the PCR tube in a thermal cycler and run the program outlined in Table 2.
Table 2. DNA Fragmentation Thermal Cycling Program
|
Temperature |
Time |
|
Heated Lid |
105°C (On) |
|
4°C |
1 min* |
|
37°C |
5–40 min** |
|
65°C |
20 min |
|
4°C |
Hold |
[Note]:
*To precisely control fragmentation and avoid over-digestion, pre-cool the thermal cycler block to 4°C before loading samples.
**For intact genomic DNA, refer to Table 3 for recommended incubation times.
Table 3. Recommended Fragmentation Times for Intact Genomic DNA
|
Target Insert Size (Peak) |
Fragmentation Time |
|
2000 bp |
5 min |
|
1000 bp |
10 min |
|
600 bp |
15 min |
|
400 bp |
20 min |
|
250 bp |
25 min |
|
150 bp |
30 min |
[Note]: For fragmented or low-integrity FFPE DNA, a 5–15 minute incubation at 37°C is recommended; optimization may be required.
5. Purify the fragmented DNA using Hieff NGSTM DNA Selection Beads (Cat#12601), AMPure XP Beads (Cat#A63880), or equivalent products. Purified DNA can be stored at –20°C for downstream applications.
Documents:
Safety Data Sheet
Manuals
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FAQ
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