Isolate and extract high quality & quantity miRNA in 30 minutes
Two column kit- eliminate large RNA on the first column and capture microRNA on a second column
Elute microRNA in 25 µL ready for miRNA profiling
Process a wide range of samples- cell, tissue, bacteria, bodily fluids, etc.
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
This kit provides a rapid method for the isolation and purification of small RNA molecules (< 200 nt) from cultured animal cells, small tissue samples, bacterial cells, plants, and blood.
Two columns are provided with this kit. The first column captures large RNA, while the small RNA are captured on a second column and are eluted concentrated in 25 µL of nuclease-free water.
The small RNA can be used in various downstream applications relating to miRNA profiling, gene regulation, and functional analysis. The eluted RNA is ready for RT-qPCR, microarrays, and NGS applications.
Background
These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. miRNAs and siRNAs are typically 20-25 nucleotides long and regulate gene expression by binding to mRNA molecules and affecting their stability or translation.
Maximum Amount of Starting Material: Animal Cells Animal Tissues Bacteria Plant Tissues Blood
3 x 106 cells 5-25 mg 1 x 109 cells 50 mg 100 μL
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
Component
Cat. 21300 (25 preps)
Buffer RL
40 mL
Wash Solution A
38 mL
Elution Solution A
6 mL
Large RNA Removal Column
25
microRNA Enrichment Column
25
Collection Tube
50
Elution Tubes (1.7 mL)
50
Product Insert
1
Other Products
DBCO-PEG9-amine TFA salt
Product Info
Document
Product Info
DBCO-PEG9-amine is a PEG linker which contains DBCO and amine moieties. The DBCO group is commonly used in copper-free Click Chemistry reactions. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The hydrophilic PEG spacer increases the water solubility of the compound. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
DBCO-PEG9-amine is a PEG linker which contains DBCO and amine moieties. The DBCO group is commonly used in copper-free Click Chemistry reactions. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The hydrophilic PEG spacer increases the water solubility of the compound. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.
Gel images of different ranges of size selection. Sheared human genomic DNA was used as input.
.
DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. 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 technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.
Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.
DNA size selection with dual clean-ups.
.
A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.
DNA size selection with single clean-up for >5 kb and >10 kb DNA.
.
Features of DNA size selection and library size selection
High specificity and high recovery of size selection
11 selection ranges are available, including 5 ranges for NGS library size selection
50-100 bp
100-200 bp
200-500 bp
250-350 bp: ideal for illumina PE100 sequencing
300-450 bp: ideal for illumina PE150 sequencing
450-750 bp: ideal for illumina PE300 sequencing
500-1000 bp
1-3 kb
1-5 kb
>5 kb: ideal for long-read sequencing
>10 kb: ideal for long-read sequencing
Fast and simple
20-min protocol
No gel purification required
No columns required
No centrifugation required
Efficient removal of contaminants and unwanted components
The HiPure FastFilter Plasmid DNA Maxi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes a special filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.
Details
Specifications
Features
Specifications
Main Functions
Isolation up to 1mg plasmid DNA from 200ml bacterial culture
Applications
Enzyme digestion, sequencing, PCR and labeling, etc.
Purification method
Maxi spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Conventional plasmid, plasmid less than 30KB
Sample amount
100-200ml
Yield
0.4-1mg
Elution volume
≥500μl
Time per run
≤60 minutes
Liquid carrying volume per column
20ml
Binding yield of column
1mg
Principle
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
Fast – it takes only 15-60 minutes to complete the isolation
Economy – high cost performance
Kit Contents
Contents
P101402
P101403
Purification Times
10 Preps
50 Preps
RNase A
20 mg
40 mg
Buffer P1
140 ml
2 x 350 ml
Buffer P2
140 ml
2 x 350 ml
Buffer LEN3
70 ml
350 ml
Buffer GBT
120 ml
550 ml
Buffer PW1
60 ml
300 ml
Buffer PW2*
50 ml
4 x 100 ml
Elution Buffer
20 ml
120 ml
HiPure DNA Maxi Columns III
10
50
Lysate Clear Maxi Syringe
10
50
50 ml Collection Tubes
20
100
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.
The HiPure FastFilter Plasmid DNA Maxi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes a special filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.
