MagBind Size-Selective DNA Purification Kit
Allows purification and selection of DNA fragments ranging from 200 bp to 1500 bp, with the selected fragments being suitable for NGS library construction.
Product Information
Product Name | Cat.No. | Spec. | Notes |
Magnetic Bead-Based DNA Fragment Purification Kit | G3613-50T | 50 T | It can be used for purification and separation of DNA fragments ranging from 200 bp to 1500 bp in PCR and enzymatic systems, with simple and fast operation. |
Description/Introduction
MagBind Size-Selective DNA Purification Kit makes use of the high efficient adsorption of nucleic acid by superparamagnetic beads, combined with specially optimized buffer system, and can purify and sort 200 bp~1500 bp DNA fragments by adding different proportions of DNA size-selective solution. The whole operation process is simple and fast, and can be used to purify DNA from PCR system and enzyme digestion system. The DNA fragments obtained by sorting can be widely used in the construction of NGS (Next Generation Sequencing) library.
Storage and Shipping Conditions
Shipped with wet ice and stored at 2~8, valid for 12 months.
Before starting (please read carefully)
1. Please prepare 80% ethanol before use, preferably ready to use.
2. Magnetic stands are required but not supplied in this kit.
- 1. Purification steps of DNA fragments
- 1) Transfer the DNA size-selective solution from 2~8 to room temperature in advance to restore the reagent to room temperature.
- 2) Upside down the DNA size-selective solution to make the magnetic beads disperse evenly. Add 1.8× volume of DNA size-selective solution to the DNA sample, gently blow with a pipette until the magnetic beads are disperse evenly, and place at room temperature for 5 min. During this period, use the pipette to blow and mix evenly for 2~3 times.
- 3) Place the centrifuge tube on the magnetic stand for 30 seconds, and after it is clear, discard the supernatant.
- 4) Add 200 μL of 80% ethanol, remove the magnetic stand, use a pipette to blow until the magnetic beads are dispersed evenly, place the centrifuge tube on the magnetic stand for 30 s, after it is clear, discard the supernatant.
- 5) Repeat step 4.
- 6) Open the centrifuge tube lid, place 5~10 min at room temperature to make the ethanol volatilize completely (avoid excessive drying of magnetic beads, so as not to affect the yield of nucleic acid).
- 7) Remove the magnetic stand, add 20~50 μL of Buffer TE or Nuclease-free Water, gently blow with the pipette until the beads are disperse evenly, and place at room temperature for 5 min.
- 8) Move the centrifugal tube to the magnetic stand until the magnetic beads are completely adsorbed, and transfer the supernatant into a new centrifuge tube to obtain high purity DNA.
- 2. Sorting steps of DNA fragment
- 1) Transfer the DNA size-selective solution from 2~8 to room temperature in advance to restore the reagent to room temperature.
- 2) The first combination: Upside down the DNA size-selective solution to make the magnetic beads disperse evenly. According to the required fragment range, add the first dosage of DNA size-selective solution according to Table 1.
- 3) Gently blow with a pipette until the magnetic beads are disperse evenly, and place at room temperature for 5 min. During this period, use the pipette to blow and mix evenly for 2~3 times.
- 4) Place the centrifuge tube on the magnetic stand for 30 seconds, and after it is clear, transfer the supernatant to a new centrifuge tube.
Note: Sample dosage×Proportion of the first DNA size-selective solution dosage=Adding amount of DNA size-selective solution, such as 100 μL sample×0.6=60 μL DNA size-selective solution
- 5) The second combination: According to Table 1, add 0.1x sample volume of DNA size-selective solution to the above centrifuge tube.
Note: Sample dosage×Proportion of the second DNA size-selective solution dosage=Adding amount of DNA size-selective solution, such as 100 μL sample×0.6=60 μL DNA size-selective solution
- 6) Gently blow with a pipette until the magnetic beads are disperse evenly, and place at room temperature for 5 min. During this period, use the pipette to blow and mix evenly for 2~3 times.
- 7) Place the centrifuge tube on the magnetic stand for 30 seconds, and after it is clear, discard the supernatant.
- 8) Add 200 μL of 80% ethanol, remove the centrifuge tube from the magnetic stand, use a pipette to blow until the magnetic beads are dispersed evenly, place the centrifuge tube on the magnetic stand for 30 s, after it is clear, discard the supernatant.
- 9) Repeat step 8.
- 10) Open the centrifuge tube lid, place 5~10 min at room temperature to make the ethanol volatilize completely (avoid excessive drying of magnetic beads, so as not to affect the yield of nucleic acid).
- 11) Remove the magnetic stand, add 20~50 μL buffer TE or Nuclease-free Water, gently blow with the pipette until the beads are disperse evenly, and place at room temperature for 5 min.
- 12) Move the centrifugal tube to the magnetic stand until the magnetic beads are completely adsorbed, and transfer the supernatant into a new centrifuge tube to obtain high purity DNA.
Table 1: Recommended DNA size-selective solution dosage
Average length of fragments (bp) | 200~ 300 | 250~ 350 | 300~ 450 | 400~ 550 | 500~ 700 | 600~ 800 | 800~ 1000 | 1000~ 1500 |
The first DNA size-selective solution dosage | 0.9× | 0.85× | 0.8× | 0.75× | 0.7× | 0.65× | 0.6× | 0.5× |
The second DNA size-selective solution dosage | 0.1× | 0.1× | 0.1× | 0.1× | 0.1× | 0.1× | 0.1× | 0.1× |
Figure 1: Sorting results of DNA fragments
Note
1. Please read the Product Manual carefully before use.
2. If the sample size is less than 100 μL, which can be replenished to 100 μL by adding water. if the sample size is too small, the actual addition of magnetic beads will be inaccurate, which will affect the sorting effect.
3. Magnetic beads are precipitate easily, so mix thoroughly before use.
4. Magnetic bead suspension should avoid freezing during preservation.
5. Ethanol should be completely volatilized before eluting the plasmid to avoid the influence of residual ethanol on downstream experiments.
6. Please do not dry the magnetic beads for a long time, so as not to affect the elution efficiency of DNA.
7. For your safety and health, please wear a lab coat and disposable gloves.