MagPure Circulating DNA Mini Kit is designed for purification of high quality circulating DNA (cfDNA) fromcell-free body fluids (such as plasma, serum). The purified DNA is suitable for direct use in downstream applications such as PCR, real-time PCR, biochip analysis and NGS.
Specifications
| Features | Specifications |
| Main Functions | Isolation circulating DNA from 0.2-0.6ml plasma, serum, body fluids |
| Applications | qPCR, NGS, etc. |
| Purification technology | Magnetic beads technology |
| Process method | Manual or automatic |
| Sample type | Serum, plasma |
| Sample amount | 0.2-0.6ml |
| Elution volume | ≥30μl |
| Time per run | ≤50 minutes |
This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by elution buffer.
| Contents | IVD5432 |
| Purification Times | 200 |
| MagPure Particles G | 14 ml |
| Carrier RNA | 310 μg |
| Proteinase K | 180 mg |
| Protease Dissolve Buffer | 10 ml |
| Buffer MLK | 250 ml |
| Buffer MAW1 | 250 ml |
| Buffer MW2* | 2 x 50 ml |
| Elution Buffer | 60 ml |
| Contents | IVD5432-F-96A | IVD5432-F-96B | IVD5432-F-96C |
| Sample amount | 200～350μl | 400~700μl | |
| Carrier RNA | 110 μg | 110 μg | |
| Proteinase K | 50 mg | 100 mg | |
| Protease Dissolve Buffer | 5 ml | 6 ml | |
| Elution Buffer | 15 ml | 15 ml | |
| Tip | 1 | 1 | |
| Sample Plate A | 500μl Buffer MLK | 500μl Buffer MLK | |
| Sample Plate B | / | 500μl Buffer MLK | |
| Wash Plate 1 | 700μl Buffer MAW1 | 700μl Buffer MAW1 | |
| Wash Plate 2 | 25μl Buffer MPG2700μl Buffer MW2 | 25μl Buffer MPG2700μl Buffer MW2 | |
| Wash Plate 3 | 700μl Buffer MW2 | 700μl Buffer MW2 | |
| Elution Plate | / | / |
| Contents | IVD5432-TL-06A | IVD5432-TL-06B | IVD5432-TL-06C |
| Sample amount | 300~350μl | 600~700μl | 900~1050μL |
| Carrier RNA | 310 μg | 310 μg | 310 μg |
| Proteinase K | 50 mg | 100 mg | 150 mg |
| Protease Dissolve Buffer | 6 ml | 6 ml | 10 ml |
| Elution Buffer | 15 ml | 15 ml | 15 ml |
| DA-Tip | 12 | 12 | 12 |
| Row 1/7 | 600μl Buffer MLK | 600μl Buffer MLK | 600μl Buffer MLK |
| Row 2/8 | / | 600μl Buffer MLK | 600μl Buffer MLK |
| Row 3/9 | 600μl Buffer MAW1 | 600μl Buffer MAW1 | 600μl Buffer MLK |
| Row 4/10 | 20μl Buffer MPG2600μl Buffer MW2 | 20μl Buffer MPG2600μl Buffer MW2 | 30μl Buffer MPG2900μl Buffer MAW1 |
| Row 5/11 | 600μl Buffer MW2 | 600μl Buffer MW2 | 900μl Buffer MW2 |
| Row 6/12 | / | / | / |
Storage and Stability
Proteinase K, Carrier RNA and MagPure Particles G should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. Theremaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least18 months under these conditions.The entire kit can be stored at 2–8°C, but in this case buffers should beredissolved before use. Make sure that all buffers are at room temperature when used.
MagPure Circulating DNA Mini Kit is designed for purification of high quality circulating DNA (cfDNA) fromcell-free body fluids (such as plasma, serum). The purified DNA is suitable for direct use in downstream applications such as PCR, real-time PCR, biochip analysis and NGS.
Details
.
Library size selection is an enrichment of a specific range of library sizes for NGS library preparations. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.
There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.
Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.
The magnetic beads, or SPRI (Solid Phase Reversible Immobilization) beads, is well used for the purification of DNA due to their reversible DNA binding. The NGS library can be size-selected by the magnetic beads or SPRI beads. The properties of the magnetic beads can be changed for a specific range of DNA binding. The contaminants and other unwanted components in the libraries can also be removed during size selection.
Specific ranges of NGS libraries can be selected using magnetic beads with different buffer compositions. The first DNA-beads binding step, also called the right-side clean-up, removes large NGS library fragments. The large NGS library fragments that bind to the beads are discarded with the beads pellet. The desired NGS library fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-DNA binding, also called the left-side clean-up. After the rinsing step, the NGS library fragments with the dual selection are eluted in water or an appropriate buffer. The magnetic beads method has great advantages over time-consuming column purification and tedious gel-based purification.
.
Library size selection for long-read sequencing only requires a single clean-up. In this case, only the large library fragments are bound to the beads, while other small library fragments are discarded with the supernatant. The selected larger library fragments are eluted in water or an appropriate buffer after the rinsing step.
.
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from FFPE tissue and section samples (with DNase) |
| Applications | RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology, DNase |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | FFPE tissue sample |
| Sample amount | 6mg |
| Yield | 20μg |
| Elution volume | ≥10μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer.
Advantages
Kit Contents
| Contents | R414402 | D414403 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Micro Columns | 50 | 250 |
| 2ml Collection Tubes | 50 | 250 |
| Buffer DPS | 60 ml | 250 ml |
| Buffer FRL | 15 ml | 60 ml |
| Buffer RLC | 15 ml | 60 ml |
| Buffer RWC* | 10 ml | 50 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| DNase I | 600 µl | 5 x 600 µl |
| DNase Booster Buffer | 1.5 ml | 6 ml |
| Protease Dissolve Buffer | 1.8 ml | 10 ml |
| Proteinase K | 24 mg | 120 mg |
| RNase Free Water | 10 ml | 20 ml |
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Experiment Data
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.