Compatible with mass spectrometry and NMR spectroscopy
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
This kit provides a fast and simple procedure for the isolation of total proteins from tissue, bacteria, yeast or mammalian cells without the use of SDS, Triton® X-100 and other detergents. Detergents are extensively used to prepare protein samples; however, these detergents have undesirable effects on downstream analysis. These effects include extraneous peaks in mass spectrometry, artifacts with chromatography and electrophoresis, interference with microinjection into cells and interference with protein immunization.
The Detergent-Free Total Protein Isolation Kit maintains high protein recovery and yields proteins that are 100% detergent-free. Purification is based on using Norgen’s proprietary resin together with Lysis Solution, followed by protein filtration using a filter column (provided). The purified proteins can be used in a number of downstream applications including mass spectrometry, SDS-PAGE, isoelectric focusing, NMR spectroscopy and more.
Each Lysis Tube is able to process up to 50 mg of tissue, 1010 bacterial cells, 109 yeast cells or 107 mammalian cells. Preparation time for 12 samples is less than 10 minutes.
Maximum Amount Of Starting Material: Tissues Animal Cells Bacteria Yeast
50 mg 1 x 107 cells 1 x 1010 cells 1 x 109 cells
Time to Process 12 Samples
Less than 10 minutes
Storage Conditions and Product Stability The Lysis Solution should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.
Component
Cat. 30300 (50 preps)
Lysis Solution
4 mL
Lysis Tubes
25
Filter Columns
25
Elution Tubes (1.7 mL)
25
Product Insert
1
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
1.Brushless frequency motor,in great torque, free maintenance, no powder pollution, quick in speed up and down.
2.Imported compressors fluorine free,double cycle cooling,cold and hot alternating easily, free environment pollution and precise in temperature control.
3. Microprocessor control, digital display which indicates the speed, time, RCF in operation, speed raising and reducing quickly, operate simply. Running parameter can be edited.
4. 10 kinds of program stored in the memory, 10 kinds of accelerating and decelerating speed for your choice.
5. Automatically electric lid lock, super speed, over temperature protection and imbalance protection. The centrifuge body is made of high quality steel, safe and reliable.
6. Rotor is connected to spindle by specialized taper sleeve, loading simple and quick, no direction.
7. There are several kinds of rotors for your choice, adapters can be designed by experimental requirements.
8. 3 tiers protection steel cover and get the ideal centrifugation result.
PACE 2.0 Genotyping Master Mix ensures an unrivalled signal-to-noise ratio and produces tight data clusters, even when working with high-throughput, crude DNA preps, resulting in consistently exceptional performance. Efficiently streamline your workflow and reduce costs without compromising the quality of your results.
PACE 2.0 Genotyping Master Mix is an ideal solution for challenging starting material. PACE 2.0 has been specially formulated to overcome the obstacles presented by common PCR inhibitor compounds, such as phenols and tannins. Even notoriously tricky samples like oil palm and conifers can still be assayed using hot shot or other crude DNA prep methods and deliver reliable and accurate data.
PACE 2.0 Genotyping Master Mix uses a novel, universal, fluorescent reporting cassette to produce machine-readable fluorescent signals corresponding to genotypes. PACE 2.0 compatible genotyping assays are comprised of two competitive allele-specific forward primers (which differ in their terminal 3’ bases and unique 5’ tail sequences) and a common, reverse primer. PACE 2.0 Genotyping Master Mix is supplied with ROX normalising dye at a range of levels to ensure compatibility with your qPCR instrument or reader.
Genotyping assay designs are available from 3CR Bioscience through our free assay-design service; once designed, users can purchase assay primers independently or through 3CR Bioscience using our partial or full-assay validation service. PACE 2.0 Genotyping Master Mix is also compatible with KASP™ and Amplifluor® marker assays.
REQUIRED COMPONENTS
qPCR machine or Thermocycler + Fluorescent plate reader
PCR plate or equivalent and appropriate optically clear seal
Template DNA
PCR-grade water
Genotyping assays
For Research and Development purposes only. Not for diagnostic use.
Legal Information KASP™ is a trademark of LGC Biosearch Technologies Amplifluor® is a registered trademark of Merck KGaA
VolumeCup
