UltraBioâ„¢ DNA Size Selection Magnetic Beads (C007B-555517)

Aladdin's UltraBio™ DNA Size Selection Magnetic Beads II are developed based on the SPRI (Solid Phase Reverse Immobilization) principle and are applicable for DNA or RNA purification and size selection during the preparation of next generation sequencing (NGS) libraries. This product can be used to recover, isolate or purify DNA or RNA fragments larger than 100bp. DNA size selection can be achieved by adjusting the amount of this product in DNA samples.Nucleic acid purification are essential for genome-related experiments, such as NGS, qPCR/ddPCR/PCR, microarray and other enzymatic reactions [1]. This product recovers DNA or RNA fragments larger than 100bp with consistency and high reproducibility. It is compatible with conventional library preparation kits and widely used in NGS and molecular diagnostics, especially in DNA or RNA purification and size selection during NGS library preparations. The yield and size distribution of NGS libraries obtained by using this product are highly consistent with those obtained from AMPure XP Beads (Beckman Coulter).This product is a salt solution containing magnetic beads, polyethylene glycol (PEG) and other substances. Under a certain concentration of salt and PEG, the conformation of some DNA molecules will change and a large number of phosphate groups will be exposed, enabling these DNA bind to the magnetic beads coated with carboxyl groups (-COOH). After the PEG and salt are removed by magnetic separation, the DNA can be eluted from the magnetic beads with appropriate solutions. Therefore, by manipulating the concentration of PEG and salt, DNA fragments of different size can be obtained [1-2]. Please refer to Figure 1 for the workflow of this product.Figure 1. The workflow of Aladdin's UltraBio™ DNA Size Selection Magnetic Beads II High SpecificityPrecautions：Although this product is similar to other products with similar functions in terms of usage and recovery rate, optimizations is still recommended for a particular experiment.This product is very similar to Mag™ DNA Size Selection Magnetic Beads (, D0038), but there are slight differences in product characteristics and specific usage. Please do not misuse these two products and their instructions.Do not freeze this product or centrifuge this product at high-speed.To ensure the recovery of DNA, the magnetic beads must be equilibrated to room temperature before use.This product must be well mixed before use by vortex or inverting the tube properly.The 80% (v/v) ethanol is recommended to be prepared fresh. Otherwise the recovery efficiency may decrease due to the evaporation of ethanol.The initial volume of the DNA sample must be greater than 100μl, if not, make up to 100μl with ultrapure water. If the sample volume is too small, the incubation of the beads and the sample will be uneven and the pipetting error will increase, thereby affecting the accuracy of size selection.Do not dry the beads for too long before elution. Excessive drying will lead to irreversible aggregation of magnetic beads and thus the reduction of elution efficiency.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1. Preparation of magnetic beadsa. Remove the magnetic beads from the 4℃ refrigerator, vortex or invert the tube upside down properly to resuspend the beads thoroughly. Take the required amount of beads and equilibrate to room temperature.b. Prepare the 80% (v/v) ethanol right before use. For example, mix 8ml of absolute ethanol and 2ml of ultrapure or ddH2O to obtain 10ml of 80% (v/v) ethanol.Note: Because the volume of ethanol and water will change after mixing, please do not measure 8ml of ethanol and add ultra-pure water or ddH2O to make up the volume to 10ml.2. Single-sided size selectionThe single-sided size selection method is mainly used to remove short DNA fragments, especially those shorter than 100bp or 200bp, such as primers or adaptor dimers, and to remove enzymes, dNTPs and salt solutions. Generally, the higher the volume ratio of magnetic beads, the higher the binding rate of short DNA fragments to magnetic beads. Therefore, the amount of magnetic beads can be adjusted to remove short DNA fragments of different length.Size-selection of DNA Bead Volumes ≥1000bp 0.6X ≥300bp 1.0X ≥200bp 1.2-1.5X ≥100bp 2.0-3.0X Note 1: The amounts of beads in the table above are for reference, and can be adjusted according to the actual test results. For example, if the volume of DNA sample is 100μl, and if a DNA fragment ≥1000bp needs to be purified, the amount of beads is 0.6X= 0.6×100μl = 60μl.Note 2: The length range here refers to the majority of the DNA fragments. A small amount of DNA fragments outside the range may still remain.a. Gently vortex or invert the tube several times to resuspend the beads thoroughly. Add an appropriate volume of beads to the DNA solution according to the table above.b. Mix thoroughly by vortex or pipetting up and down for 10 times. Incubate at room temperature for 5 minutes.c. Spin the tube briefly and place it on magnetic stand for 5 minutes. After the solution is clear, aspirate the supernatant carefully.d. Keep the tube in the magnetic stand, add 200μL of freshly prepared 80% (v/v) ethanol without disturbing the beads, and incubate at room temperature for 30 seconds. Aspirate and discard the ethanol.e. Repeat step d once.f. Keep the tube in the magnetic stand, air dry the beads with the lid open until cracks just appear (5-10 minutes).Note: Do not over-dry the beads, which may result in lower elution efficiency.g. Remove the tube from the magnetic stand. Add an appropriate amount of ddH2O (≥20 μL) and mix thoroughly by vortex or pipetting.h. Incubate at room temperature for 5 minutes.i. Spin the tube briefly and place it on the magnetic stand for 5 minutes. After the solution is clear, transfer 20μL of the supernatant to a new microcentrifuge tube. Please refer to Figure 2 for the performance of this product in DNA Sorting.Figure 2. Single-sided size selection of DNA using DNA Size Selection Magnetic Beads II . DNA in 100μl solution (2μg DNA, 100-10000bp) was purified by adding 0.6X-3X volumes of beads, respectively. A small amount of the purified products was mixed with DNA loading buffer (6X) ( DNA Size Selection Magnetic Beads II . DNA in 100μl solution (1μg DNA, 100-10000bp) was purified according to this protocol. A small amount of purified products was mixed with DNA loading buffer (6X) (, D0071) and examined by 1% agarose gel electrophoresis. This figure is for reference only, which may vary due to different experimental conditions.