Water-soluble, substrate for sortase mediated labeling of proteins. Sortase catalyzes a transpeptidase reaction between a specific internal sequence of a protein and an amine group present on the N-terminus of triglycine recently has become an area of great interest. This method of labeling proteins has been denoted as “Sortagging”. Proteins conjugated to DBCO-Gly-Gly-Gly can be further modified with azide-containing molecules creating site-specific protein conjugates. Examples of creating protein conjugates using sortagging include site-specifically PEGylating proteins,1 site-specific protein-lipid conjugates,2 and constructing peptides and glycosylphosphatidylinositol chimeras.3 Sortase has also been used in peptide synthesis to cyclize peptides to create macrocyclic peptides, glycopeptides4 and protein−protein conjugates.
Detail
Water-soluble, substrate for sortase mediated labeling of proteins. Sortase catalyzes a transpeptidase reaction between a specific internal sequence of a protein and an amine group present on the N-terminus of triglycine recently has become an area of great interest. This method of labeling proteins has been denoted as “Sortagging”. Proteins conjugated to DBCO-Gly-Gly-Gly can be further modified with azide-containing molecules creating site-specific protein conjugates. Examples of creating protein conjugates using sortagging include site-specifically PEGylating proteins,1 site-specific protein-lipid conjugates,2 and constructing peptides and glycosylphosphatidylinositol chimeras.3 Sortase has also been used in peptide synthesis to cyclize peptides to create macrocyclic peptides, glycopeptides4 and protein−protein conjugates.
Other Products
Bis-propargyl-PEG10
Product Info
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Product Info
Bis-propargyl-PEG10 is reactive with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry. By introducing PEG units into the molecule, the solubility of the compound in aqueous environment can be improved. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
Bis-propargyl-PEG10 is reactive with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry. By introducing PEG units into the molecule, the solubility of the compound in aqueous environment can be improved. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
The HiPure Minipreps system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Miniprep 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 30μg endotoxin-free plasmid DNA from 1-5ml bacterial culture using 96-well bind plate
Applications
Cell transfection, animal injection, etc.
Purification method
96 well plate
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Low copy plasmid vector
Sample amount
1-5ml LB(x96)
Yield
30μg
Elution volume
≥75μl
Time per run
≤60 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
1. High throughput: processing 96 samples at once; 2. High yield: The purified plasmid yield can reach up to 30μg, which is greater than the yield of the magnetic bead method kit; 3. Detoxification: endotoxin content <0.1EU/μg; 4. Fast: The entire extraction can be completed in 60 minutes without the need for time-consuming alcohol precipitation;
Kit Contents
Contents
P115701
P115702
P115703
Purification Times
1 x 96 Preps
4 x 96 Preps
20 x 96 Preps
RNase A
10 mg
20 mg
100 mg
Buffer P1
30 ml
120 ml
550 ml
Buffer P2
30 ml
120 ml
550 ml
Buffer LEN3
15 ml
60 ml
300 ml
Buffer LN4
80 ml
270 ml
2 x 700 ml
Buffer LN5
40 ml
220 ml
1100 ml
Buffer PW1
40 ml
220 ml
1100 ml
Buffer PW2
25 ml
100 ml
2 x 200 ml
Elution Buffer
30 ml
60 ml
250 ml
Lysate Clear Plate
1
4
20
HiPure DNA Plate
1
4
20
1.6ml Collection Plate
2
8
40
0.5ml Collection Plate
1
4
20
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 Minipreps system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Miniprep 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.
Cat.# 20110S, 20110L: Size range >5 kb (ideal for long-read sequencing size selection)
Product Info
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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.
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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