[DL5000] FluoroDye™ DNA Fluorescent Loading Dye (Green, 6X), 1 ml
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FluoroDye™ DNA Fluorescent Loading Dye is a ready-to-use 6X DNA loading dye designed for fast qualitative electrophoresis analysis. Containing sensitive fluorescent dye with high specific affinity towards double stranded DNA (dsDNA), the FluoroDye™ Fluorescent DNA Loading Dye has negligible background and renders destaining process unnecessary. The FluoroDye™ DNA Fluorescent Loading Dye allows the user to immediately visualize electrophoresis result upon completion or to monitor the electrophoresis in real time. FluoroDye™ DNA Fluorescent Loading Dye is compatible with both the conventional UV gel-illuminating system as well as the less harmful long wavelength blue light illumination system. FluoroDye™ emission as bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.
Detail
Description
FluoroDye™ DNA Fluorescent Loading Dye is a ready-to-use 6X DNA loading dye designed for fast qualitative electrophoresis analysis. Containing sensitive fluorescent dye with high specific affinity towards double stranded DNA (dsDNA), the FluoroDye™ Fluorescent DNA Loading Dye has negligible background and renders destaining process unnecessary. The FluoroDye™ DNA Fluorescent Loading Dye allows the user to immediately visualize electrophoresis result upon completion or to monitor the electrophoresis in real time. FluoroDye™ DNA Fluorescent Loading Dye is compatible with both the conventional UV gel-illuminating system as well as the less harmful long wavelength blue light illumination system. FluoroDye™ emission as bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.
Features:
Excellent for premix with DNA samples
Sensitivity: 0.14 ng (DNA)
A safer alternative to EtBr
Compatibility: suitable to blue or UV light
Increased cloning efficiency (blue light)
Composition
FluoroDye™ DNA Fluorescent Loading Dye is stored in 6X concentration in 60% glycerol and buffered with Tris-HCl and EDTA, containing Bromophenol blue, Xylene cyanol FF and Orange G as tracking dyes.
Storage
Protected from light -20°C for 24 months
Other Products
ITS2 Library Preparation Kit for Illumina
Product Info
Document
Product Info
Overview
Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
Simple and quick workflow: library could be prepared in less than 4 hours
Component of Norgen’s metagenomics workflow
A single NGS run can be prepared with up to 384 unique dual-index libraries
The ITS2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the fungal target ITS2 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 ITS2 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 ITS2 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 ITS2 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.
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.
Details
Specifications
Features
Specifications
Main Functions
Isolation up to 70μg plasmid DNA from 5-15ml 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 plasmid : 5-10ml culture medium
Yield
20-80µg
Elution volume
≥75μl
Time per run
Complete 1-24 samples in 30 minutes
Liquid carrying volume per column
800µl
Binding yield of column
70µg
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 2 minutes to obtain supernatant by optimized solution (10 minutes for other brands)
High yield – up to 70µg plasmid can be binded in one column
Kit Contents
Contents
P100202
P100203
Purification Times
50 Preps
250 Preps
RNase A
5 mg
20 mg
Buffer P1
30 ml
140 ml
Buffer P2
30 ml
140 ml
Buffer P3
40 ml
200 ml
Buffer PW1
30 ml
140 ml
Buffer PW2*
12 ml
50 ml
Elution Buffer
15 ml
30 ml
HiPure DNA Mini Columns III
50
250 ml
2 ml Collection Tubes
50
250 ml
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.