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
D3142 HiPure Soil DNA Kit
Product Info
Document
Product Info
Introduction
Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:
1. Humic acid pollution. The soil, especially in forests and grasslands, is rich in humic acids. Humic acid is a series of organic molecules, some of which are very similar to nucleic acid molecules and difficult to remove during purification. Trace amounts of humic acid pollution can lead to downstream applications such as PCR and enzyme digestion failure.
2. Lysis method. Soil samples contain various microorganisms, such as bacteria and fungi. Gram positive bacteria and fungi both contain very thick bacterial walls, and effectively breaking down the cell walls of these microorganisms is crucial for extracting high-yield metagenomic DNA. Due to the complexity of soil samples, it is not feasible to use enzymatic methods (such as lysozyme, wall breaking enzyme, snail enzyme) or liquid nitrogen grinding, as the soil contains various metalions or inhibitory factors that inactive the digestive enzymes, or the presence of sand particles in the soil makes liquid nitrogen grinding difficult.
3. The DNA yield is difficult to control. Soil samples would have significant changes in the number and variety of microorganisms due to fertility, inferiority, high moisture content, dryness, or depth of sampling. In a small range of soil samples, the DNA content often varies by thousands of times. In addition, certain chemical components in soil, such as heavy metal salts and clay substances, can cause a decrease in DNA yield.
Magen’s HiPure Soil DNA Kits are currently the most optimized kit for soil DNA extraction. The kit adopts glass bead grinding method and thermal shock chemical wall breaking method, which can be carried out in the point vortex instrument without special bead grinding instrument, and is suitable for a wide range of laboratories. The Absorber Solution in the reagent kit is a humic acid adsorbent exclusively developed by Magen Company, which can efficiently remove various humic acid pollutants. In addition, an alcohol-free silica gel column purification method is also used to efficiently remove various soluble metal salts and other soluble inhibitory factors from the soil. The kit has successfully extracted from the following soil (partially based on customer feedback): soil from forests in nature reserves (30 to 40 years old forest soil with a surface layer of 30-50cm deciduous layer), mangrove soil, grasslands, farmland, seabed mud, sludge, mineral area soil, organic matter contaminated soil, pond mud, garbage mud, air conditioning pipeline deposits, etc.
This product allows rapid and reliable isolation of high-quality genomic DNA from various soil samples. Up to 500 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatilityof spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.
Details
Specifications
Features
Specifications
Main Functions
Isolation DNA from 200-500mg soil sample
Applications
PCR, southern blot and enzyme digestion, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Soil
Sample amount
200-500mg
Elution volume
≥30μl
Time per run
≤60 minutes
Liquid carrying volume per column
800μl
Binding yield of column
100μg
Principle
Soil sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. humic acid,proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.
Advantages
Fast – several samples can be extracted in 40 minutes (after digestion)
High purity – purified DNA can be directly used in various downstream applications
Good repeatability – silica technology can obtain ideal results every time
High recovery – DNA can be recovered at the level of PG
Kit Contents
Contents
D314202
D314203
Purification Times
50 Preps
250 Preps
Hipure DNA Mini Columns II
50
250
2ml Collection Tubes
50
250
2ml Bead Tubes
50
250
Buffer SOL
60 ml
250 ml
Buffer SDS
5 ml
20 ml
Buffer PS
10 ml
50 ml
Absorber Solution
10 ml
50 ml
Buffer GWP
40 ml
220 ml
Buffer DW1
30 ml
150 ml
Buffer GW2*
20 ml
2 x 50 ml
Buffer AE
15 ml
30 ml
Storage and Stability
Absorber Solution should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
Experiment Data
Document
Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:
Isolate all sizes of circulating and exosomal RNA, including microRNA
Versatile plasma/serum input ranges
No phenol extractions
No carrier RNA
Bind and elute all RNA irrespective of size or GC content, without bias
Concentrate circulating RNA and exosomal RNA into a flexible elution volume
High quality, purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Compatible with Streck Cell-Free RNA BCT® Tubes
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
Background
Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as plasma or serum.
Plasma/Serum RNA Purification Mini Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 µL to 200 µL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 µL to 25 µL.
Plasma/Serum RNA Purification Midi Kit
This utilizes a two column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 µL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
Plasma/Serum RNA Purification Maxi Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
*This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.
**Please check page 7 for Average Plasma/Serum Yields and Common RNA Quantification Methods.
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
DBCO-NHCO-PEG4-acid is an analog of DBCO-Acid with a hydrophilic PEG spacer arm, which improves water solubility. This reagent is a non-activated building block with enhanced solubility in aqueous media and can thus be used to derivatize amine-containing molecule through a stable amide bond. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
DBCO-NHCO-PEG4-acid is an analog of DBCO-Acid with a hydrophilic PEG spacer arm, which improves water solubility. This reagent is a non-activated building block with enhanced solubility in aqueous media and can thus be used to derivatize amine-containing molecule through a stable amide bond. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.