The ITS1 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the fungal target ITS1 libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XPMagneticBeads(supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal/swab, plasma/serum, tongue swab, gum swab, and others.
The ITS1 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the ITS1 region of the fungal DNA.The post-PCR reaction is then cleaned up using AMPure XPbeads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XPbeads. The libraries are then ready for quantification, pooling and sequencing.
Storage Conditions and Product Stability Norgen’s ITS1 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.
All kit components will remain stable for at least 1 year when stored at the specified storage conditions.
The Plant RNA Kit (NA2014-01) is optimized for purification of total RNA from a wide variety of plant samples which rich in polysaccharides and phenol. It comes with proprietary magnetic beads and a specially optimized buffer. Toxic solvents such as phenol, chloroform are not needed in the process. The eluted RNA is high-quality and is ready for all downstream applications, such as RT-PCR, Northern blotting, differential display. The procedures can be fully automated on the magnetic particle processor instrument and ease of use.
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.
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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.
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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.
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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
Collect from nasal, buccal, saliva, fecal, skin, surfaces, and more
Completely stabilize microbiota profiles from the point-of-collection
Minimize bias in data analysis: NGS (Metagenomics/Microbiome), PCR and microarrays
No need to immediately process samples: ship and store at ambient temperatures
DNA preservation at room temperature over 2 years
Renders sample non-infectious for safe and easy shipping
Compatible with DNA isolation kits and protocols: Optimized isolation procedure using Norgen’s Microbiome DNA Isolation Kit
Norgen’s Swab Collection and DNA Preservation System is designed for collection, ambient storage and transport of DNA from samples collected using a swab, including nasal, buccal, saliva, fecal, skin, surfaces, etc. The Swab Collection and DNA Preservation System contains Norgen’s Swab Preservative in a liquid format. The user simply collects the specimen using the provided swab, and then transfers the swab into the Swab Preservative. The Swab Preservative prevents the growth of Gram-negative and Gram-positive bacteria and fungi, and also inactivates viruses allowing the resulting non-infectious samples to be handled and shipped safely. In addition, the Swab Preservative eliminates the need to immediately process or freeze samples and allows the samples to be shipped to centralized testing facilities at ambient temperatures. The components of the Swab Preservative allow DNA samples to be stored at room temperature for over 2 years.
DNA Isolation from Preservative
Swab samples collected and preserved using Norgen’s Swab Collection and DNA Preservation System are compatible with most DNA isolation methods. Samples stored in the tubes have been used successfully with most of Norgen Biotek’s DNA isolation kits and reagents. An optimized isolation procedure is available as part of Norgen’s Microbiome DNA Isolation Kit (Cat. 64100).
When stored at room temperature, unused Norgen Swab Preservative Tubes are stable through to the collect-before date without any reduction in performance. Please see the device insert or tube label for collect-before date.
