Quantify RNA of a wide spectrum of concentrations, including the lower ng per µL and pg per µL range
Compatible with any Real-Time PCR system
RNA is accurately quantified using a standard curve constructed from the provided RNA standard
Norgen’s Low Abundance RNA Quantification Kit offers a PCR-based detection procedure to quantify RNA of a wide spectrum of concentrations, including the lower ng per µL and pg per µL range. The kit has two main enzymatic components – reverse transcription using Norgen’s microScript Reverse Transcription system and PCR Master Mix used in conjunction with a specially formulized primer mixture, to amplify human RNA from different types of inputs (such as various liquid biopsies). The kit is compatible with any Real-Time PCR system with the addition of fluorescent nucleic acid stains such as SYBR Green. The unknown RNA is accurately quantified by using a standard curve constructed from the provided RNA Standard.
Storage Conditions Upon receipt, store Norgen’s Low Abundance RNA Quantification Kit at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.
Gel images of different ranges of library size selection. Sheared human genomic DNA was used as input.
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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.
NGS library size selection with dual clean-ups.
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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.
NGS library size selection with single clean-up for >5 kb and >10 kb libraries.
The HiPure Mini system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Plasmid Mini Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of tris buffer or water.
Compared to other domestic products, Magen was the first to solve the stability problem of the column. Many other brands have unstable extraction concentrations, and the longer the time, the more unstable the column becomes. However, in our test of Magen kit, the quality and yield of plasmid extraction still remain stable after 5 years’ storage. For different customers, our kits can be customized. For example, Classic type is suitable for customers with low copy or unclear plasmid types. The rapid type is suitable for customers with high copy plasmids. Compared to many other brands, the plasmid DNA extracted by Magen has a longer preservation time and more thoroughly RNA removal.
Details
Specifications
Features
Specifications
Main Functions
Isolation up to 35μg plasmid DNA from 1-5ml bacterial culture
Applications
Enzyme digestion, sequencing, PCR, cloning, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Conventional plasmid, plasmid less than 30KB
Sample amount
High copy plasmid: 1-5ml culture mediumLow copy number plasmid : 5-10ml culture medium
Yield
5-35µg
Elution volume
≥30μl
Time per run
Complete 1-24 samples in 30 minutes
Liquid carrying volume per column
800µl
Binding yield of column
35µg
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 1 minute to obtain supernatant by optimized solution (10 minutes for other brands)
High yield – up to 35µg plasmid can be binded in one column
Kit Contents
Contents
P100102
P100103
Purification Times
100 Preps
250 Preps
RNase A
5 mg
10 mg
Buffer P1
30 ml
80 ml
Buffer P2
30 ml
80 ml
Buffer P3
40 ml
100 ml
Buffer PW1
60 ml
140 ml
Buffer PW2*
20 ml
50 ml
Elution Buffer
15 ml
30 ml
HiPure DNA Mini Columns II
100
250
2 ml Collection Tubes
100
250
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 Mini system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Plasmid Mini Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of tris buffer or water.
