ArcticZymes Proteinase is an unspecific endopeptidase originating from an Arctic marine microbial source. It has broad substrate specificity and is easy to inactivate after use.
Histones and other proteins are known to protect nucleic acids from interacting optimally with other DNA binding proteins and enzymes. ArcticZymes Proteinase is ideally suited for transforming chromatin and other dense nucleic acids to naked DNA. The enzyme is easy to heat-inactivate. This allows thermal inactivation at temperatures allowing RNA integrity as well as avoiding dissociation of dsDNA.
Detail
OverView
ArcticZymes Proteinase is an unspecific endopeptidase originating from an Arctic marine microbial source. It has broad substrate specificity and is easy to inactivate after use.
Histones and other proteins are known to protect nucleic acids from interacting optimally with other DNA binding proteins and enzymes. ArcticZymes Proteinase is ideally suited for transforming chromatin and other dense nucleic acids to naked DNA. The enzyme is easy to heat-inactivate. This allows thermal inactivation at temperatures allowing RNA integrity as well as avoiding dissociation of dsDNA.
Key Features
Easy to inactivate after use: 15 – 30 min at 55 – 60°C
Broad substrate specificity
Active at high salt concentrations
Compatible with downstream DNA analyses
Available in Glycerol FREE formulation
Applications
Lysis and releasing nucleic acid from single-cells and small tissue samples
Optimization of DNA/RNA yields from nucleic acids when added to extraction procedures
Figures
Other Products
Cat.# 20102S, 20102L: Size range 100-200 bp
Product Info
Document
Product Info
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
Neisseria gonorrhoeae is a Gram-negative coccus of the Neisseria genus. N. gonorrhoeae is usually seen in pairs infecting human cells. It has a circular DNA genome of approximately 1Mbp encoding over 2000 genes. N. gonorrhoeae is transmitted by sexual contact and usually causes infection in cells of the mucous membrane of the male urethra or the endocervix and urethra in females. During infection, polysaccharides are released from the bacteria that stimulate host cell production of tumour necrosis factors that cause an inflammatory response.
There is no vaccine against N. gonorrhoeae infection and antibiotic resistance is beginning to increase, therefore treatment includes a course of antibiotics that will be effective against resistant strains. Complications in males caused by the infection can result in prostatitis or orchitis if the bacteria spread. In females, invasion of the fallopian tubes or ovaries can result in salpingitis or ovaritis respectively, with any of these infections possibly resulting in sterility.
Storage Conditions and Product Stability All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
Sequentially isolate and purify total RNA and DNA from a single sample
Two column system: one for DNA and one for RNA
The RNA column is for the purification of total RNA including microRNA
No need to split the lysate, or to use phenol or precipitation methods
Rapid and efficient spin column procedure – it takes only 30 minutes
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
These kits provide a rapid method for the isolation and purification of total RNA and DNA sequentially from a single sample of cultured animal cells and tissues, blood, bacteria, yeast, or fungi. The lysate is passed over two columns: 1) a DNA column and 2) an RNA column. Total RNA of all sizes is purified, including microRNA. Both DNA and RNA are of the highest purity and yield.
These kits are ideal for researchers who are interested in studying the genome and transcriptome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/DNA Purification Kits are especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as they eliminate the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and DNA are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications
RNA/DNA Purification Kit (Spin Column)
Maximum column binding capacity of 50 μg for RNA and 20 μg for DNA.
RNA/DNA Purification Micro Kit (Micro)
The purified RNA and DNA fractions can be eluted in as little as 20 μL. Ideal for cell number inputs of 500,000 and as little as 5 cells. Maximum column binding capacity of 35 μg for RNA and 10 μg for DNA.
Maximum Amount of Starting Material: Animal Cells Animal Tissues Blood Bacteria Yeast Fungi Plant Tissues
5 x 106 cells 25 mg (for most tissues)** 200 μL 1 x 109 cells 1 x 108 cells 50 mg 50 mg
Time to Complete 10 Purifications
30 minutes
Average Yield*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg)
10-15 μg RNA 2-4 μg DNA 30-35 μg RNA 4-6 μg DNA
*Average Yield will vary depending upon a number of factors including species, growth conditions used, and development stage.
**Tissue inputs of up to 40 mg may be used, however for inputs greater than the recommended 25 mg, cross-contamination of the RNA and DNA fractions is possible.
Storage Conditions and Product Stability Store Proteinase K at -20°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
