NH-bis(PEG4-Propargyl) is a bifunctional PEG compound containing two terminal alkynes that are joined together at a secondary amine. Terminal alkynes are reactive towards azides in copper (I) click chemistry to form stable triazoles with the target compound. The secondary amine joining the two arms can be used as a nucleophile such as in alkylation via reductive amination or in forming amides with carboxylic acids or activated NHS esters. The use of a central amine also allows for hydrogen bonding, further increasing this compound’s water solubility.

NH-bis(PEG4-Propargyl) is a bifunctional PEG compound containing two terminal alkynes that are joined together at a secondary amine. Terminal alkynes are reactive towards azides in copper (I) click chemistry to form stable triazoles with the target compound. The secondary amine joining the two arms can be used as a nucleophile such as in alkylation via reductive amination or in forming amides with carboxylic acids or activated NHS esters. The use of a central amine also allows for hydrogen bonding, further increasing this compound’s water solubility.

Mal-amido-PEG4-alkyne is a short PEG linker featuring a maleimide and a terminal alkyne. Maleimide is a covalent ligand used for linking thiols such as those in cysteines residues in proteins, while terminal alkynes can be used in copper (I) click chemistry with azides on target molecules.

Mal-amido-PEG4-alkyne is a short PEG linker featuring a maleimide and a terminal alkyne. Maleimide is a covalent ligand used for linking thiols such as those in cysteines residues in proteins, while terminal alkynes can be used in copper (I) click chemistry with azides on target molecules.

Introduction

The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from 1-1.5ml whole blood
Applications	qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection
Purification method	Mini spin column
Purification technology	Silica technology, acidphenol / guanidine extraction technology (MagZol pretreatment technology)
Process method	Manual (centrifugation or vacuum)
Sample type	Fresh or frozen blood, mammalian blood
Sample amount	≤1.5 ml whole blood
Yield	2-100μg
Elution volume	≥20μl
Liquid carrying volume per column	800µl
Binding yield of column	100μg

Principle

The Kit simplifies isolation of RNA from blood with a fast spin-column procedure. Red blood cells are selectively lysed and white cells collected by centrifugation. White cells are then lysed using highly denaturing conditions which immediately inactivate RNases. After homogenization using the DNA spin column, the sample is applied to the RNA column. Total RNA binds to the membrane and contaminants are washed away, leaving pure RNA to be eluted in 30–100µl RNase-free water (provided with the kit) for direct use in any downstream application.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – isolation of several samples can be completed in 20 minutes by using column purification method
• Sensitive – RNA can be purified at the level of PG

Kit Contents

Contents	R416102	R416103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns	50	250
HiPure RNA Mini Columns I	50	250
2ml Collection Tubes	100	500
10 x Buffer RBC	50 ml	3 x 100 ml
RTL Lysis Buffer	50 ml	250 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

HiPure Blood RNA Mini Kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form in the RTL Lysis Buffer. Dissolve by warming buffer to 37°C.

The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection and in vitro translation.