NGS DNA selection Beads HYD511
Product Description
NGS DNA selection Beads are prepared based on the SPRI (Solid Phase Reverse Immobilization) principle and is applicable for DNA purification and size selection during the preparation of next generation sequencing (NGS) libraries. NGS DNA selection Beads is compatible with various of DNA and RNA library prep kits and is a good alternative of AMPure beads.
Product Details
Components
|
Components |
Size-1 |
Size-2 |
Size-3 |
|
NGS DNA selection Beads |
1ml |
5ml |
60ml |
Storage
Store at 2-8℃ for 1 year.
Instructions
1. Preparation
Equilibrate the selection beads at room temperature for at least 30 min before use.
2. Size selection
The operation flow of size selection is shown in Figure 1 and the protocol is as follows.
1) Mix the beads thoroughly by vortexing or pipetting up and down every time before using.
2) Add the first round of selection beads to the sample (refer to Table 1). Mix thoroughly by vortexing or pipetting up and down at least 10 times.
3) Incubate at room temperature for 5 min.
4) Spin down the tube briefly and place it on magnetic stand. When the solution is clear (about 5 min), transfer thesupernatant to a new PCR tube.
5) Add the second round of selection beads to the sample from step 2.4 according to Table 1. Mix thoroughlybyvortexing or pipetting up and down at least 10 times.
6) Incubate at room temperature for 5 min.
7) Spin down the tube briefly and place it on magnetic stand. When the solution is clear (about 5 min), aspiratethesupernatant and discard.
8) Keep the tube in the magnetic stand and add 200 μL of freshly prepared 80% ethanol to without disturbingthebeads, incubate at room temperature for 30 sec. Aspirate the ethanol and discard.
9) Repeat step 2.8 once for a total of two washes.
10) Remove residual ethanol with 10 µL pipette tips. Keep the tube in the magnetic stand, air dry the selectionbeads with the lid open until cracks just appear (about 5 min).Note: Do not over-dry the selection beads. This may result in lower recovery DNA target.
11) Remove the tube from the magnetic stand. Add an appropriate amount of ddH2O (≥20 µL) and mix thoroughlyby vortexing or pipetting up and down at least 10 times.
12) Incubate at room temperature for 5 min.
13) Spin down the tube briefly and place it on the magnetic stand. When the solution is clear (about 5 minutes), transfer 20 μL of the supernatant to a new tube.
3. Recommended Conditions for DNA Size Selection The calf thymus DNA was fragmented by sonication to prepare a fragment of 100-1,000 bp, and two rounds of sizeselection were performed according to Table 1. The results were analyzed using Agilent 2100 Bioanalyzer (Figure2).
Table 1. Recommended condition for DNA size selection
|
Length of DNA fragment |
250-350 bp |
320-420 bp |
450-550 bp |
550-700 bp |
700-900 bp |
800-1,000 bp |
|
Ratio of Beads: DNA for the 1st Round |
0.80× |
0.70× |
0.60× |
0.55× |
0.50× |
0.45× |
|
Ratio of Beads: DNA for the 2nd Round |
0.20× |
0.20× |
0.20× |
0.15× |
0.15× |
0.15× |
Note: "×" in the table indicates the volume of sample DNA. For example, if the insert length of the library is 250 bp and the sample DNAvolume is 100 μL, the volume of magnetic beads used in the first round of sorting is 0.80×100 μL=80 μL; the volume of magnetic beadsused in the second round of sorting is 0.20× 100 μL=20 μL
Figure 2. Agilent 2100 high sensitivity DNA chip electropherogram
Notes
1. For your safety and health, please wear a lab coat and disposable gloves.
2. The magnetic beads must be equilibrated at room temperature for at least 30 min before use.
3. 80% ethanol must be prepared immediately before use, otherwise it will affect the recovery efficiency.
4. When performing length sorting, the initial sample volume must be ≥100μL. If it is insufficient, please use ultrapure water to make up. If the sample volume is too small, it will increase the pipetting error, thereby affecting the accuracy of the sorting.
5. This product is for scientific research purposes only!