Propargyl-PEG14-t-butyl ester

N-Boc-N-bis(PEG4-Propargyl) is a bifunctional PEG compound containing two terminal alkynes that are combined together at a Boc-protected secondary amine. Terminal alkynes are most commonly used in copper click chemistry with azides to form stable triazoles with the target molecule. The secondary amine may be deprotected under acidic conditions to allow for alkylation at that position, increasing the molecule’s structural complexity. The use of a central amine also allows for hydrogen bonding, further increasing this compound’s water solubility.

N-Boc-N-bis(PEG4-Propargyl) is a bifunctional PEG compound containing two terminal alkynes that are combined together at a Boc-protected secondary amine. Terminal alkynes are most commonly used in copper click chemistry with azides to form stable triazoles with the target molecule. The secondary amine may be deprotected under acidic conditions to allow for alkylation at that position, increasing the molecule’s structural complexity. The use of a central amine also allows for hydrogen bonding, further increasing this compound’s water solubility.

Introduction

Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.

The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.

Details

Specifications

Features	Specifications
Recommended application	Small amounts of nucleic acid isolation, viral nucleic acid from cell free samples
Preservation conditions	Room temperature
Stability	Up to 4 years
Filter membrane	High quality glass fiber filter GF/F, 2 layers
Membrane aperture	0.7μm
Maximum binding yield of plasmid	30 μg
Maximum yield of alcohol mediated Binding	100 μg
Single liquid carrying capacity of column	900 μl
Minimum elution volume	80 μl
Withstand centrifugal force	5,000 x g
Centrifuge	Low speed centrifuge, Swing out Rotor, can placed a height of 6.5cm square,  (height of HiPure DNA Plate & 1.6ml Collection Plate: height, 6.2cm)

Adsorption Mechanism

Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silica gel membrane, while other pollutants, mainly proteins, are removed by membrane washing.

Ordering information

CAT.No.	Product Name	Package
C13130	HiPure DNA Plate (2 x GF/F)with 1.6ml Collection Plate	10/Bag

Purchase Guide

Item No.	Product Name	Membrane type/number of layers	Collection tubes	Plasmid DNA binding capacity (Physical adsorption)	gDNA/RNA binding capacity (Alcohol-mediated adsorption)	Minimum Elution volume	Liquid volume capacity
C13010	HiPure DNA Nano Column	2 layers GF/F	2ml without cap	5μg	20μg	10μl	700μl
C13011	HiPure DNA Micro Column	3 layers GF/F	2ml without cap	10μg	50μg	15μl	700μl
C13100	HiPure DNA Mini Column I	2 layers GF/B	2ml without cap	15μg	100μg	30μl	700μl
C13110	HiPure DNA Mini Column II	4 layers GF/B	2ml without cap	35μg	200μg	50μl	800μl
C13111	HiPure RNA Mini Column	3 layers GF/B	2ml without cap	30μg	200μg	30μl	800μl
C13112	HiPure Viral Mini Column	3 layers GF/F	2ml without cap	30μg	200μg	30μl	800μl
C13113	HiPure CFDNA Mini Column	3 layers GF/F,1 layer GF/B	2ml without cap	30μg	200μg	30μl	800μl
C13120	HiPure DNA Midi Column	4 layers GF/B	15ml Centrifuge tube	125μg	1mg	500μl	4ml
C13121	HiPure DNA Midi Column III	8 layers GF/B	15ml Centrifuge tube	250μg	1mg	500μl	4ml
C13122	HiPure DNA Maxi Column	4 layers GF/B	50ml Centrifuge tube	500μg	5mg	1000μl	20ml
C13123	HiPure DNA Maxi Column III	8 layers GF/B	50ml Centrifuge tube	1mg	5mg	1000μl	20ml
C13124	HiPure DNA Maxi Column C	8 layers GF/B	50ml high speed Centrifuge tube	1mg	5mg	700μl	12ml
C13130	HiPure DNA Plate	2 layers GF/F	1.6ml Plate	30μg	100μg	80μl	900μl
C13131	HiPure gDNA Plate	2 layers GF/B	1.6ml Plate	30μg	100μg	80μl	900μl

Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.

Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.

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 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The 16S V1-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 amplicon 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 XP Magnetic Beads (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 16S V1-V2 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 V1-V2 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. 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 XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

Figure 1 / 3

Previous

Next

Click for expanded view

Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V1-V2 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 should remain stable for at least 1 year when stored at the specified storage conditions.

Step	Component	Cat. 70100 (24 preps)	Cat. 70110, 70120, 70130, 70140 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-V2 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL