Introduction

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA and miRNA from cell and tissue without MagZol reagent
Applications	RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Cell, Animal tissue, Plant tissue
Sample amount	Cells: ≤ 5 x 10^6, Animal tissue:<10mg
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Biological samples are first lysed and homogenized in a highly denaturing guanidine isothiocyanate-containing buffer, which immediately inactivates DNases and RNases to ensure isolation of intact DNA and RNA. The lysate is then passed through a Mini spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. Flow-through from the column is digested by Proteinase K in the presence of ethanol. This optimized digestion, together with the subsequent addition of further ethanol, allows appropriate binding of total RNA, including miRNA, to the column. Contaminants are efficiently washed away and high-quality RNA is eluted.

Advantages

• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – several samples can be extracted in 40 minutes
• High applicability – samples including animals, plants, bacteria, cells, etc.
• High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity
• Safe – no phenol chloroform extraction and no use of Trizol reagent

Kit Contents

Contents	R431102	R431103
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	100	2 x 250
2ml Collection Tubes	100	2 x 250
Proteinase K	48 mg	240 mg
Protease Dissolve Buffer	5 ml	15 ml
Buffer RLC	40 ml	200 ml
Buffer RWC	20 ml	80 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	60 ml

Storage and Stability

Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.

Description 

ExcelTaq™ Klen-Taq DNA Polymerase is a specially blended enzyme mix containing KlenTaq-1 DNA polymerase (a 5’-exo-minus, N-terminal deletion of Taq DNA polymerase) and a small amount of a proofreading DNA polymerase. This unique blending helps to improve the fidelity, yield and processivity of the resultant PCR process. The Klen-Taq is also highly robust, showing high tolerance of varying concentrations of Mg²⁺; it is highly thermostable and has four times the fidelity compared to Taq DNA polymerase. The ExcelTaq™ Klen-Taq DNA Polymerase is ideal for DNA amplifications 0.5-5 kb in length on genomic DNA, and up to 10 kb on less complex templates.

Features

• 5’→3′ DNA polymerase activity
• 3’→5′ exonuclease activity (proofreading)
• 4× fidelity as compared to Taq DNA polymerase
• Thermo-stable: up to 98°C during PCR denaturing step
• Robust PCR performance, resistance to variance in PCR conditions 

Storage

-20°C for 24 months 

ExcelTaq™ Klen-Taq DNA Polymerase is a specially blended enzyme mix containing KlenTaq-1 DNA polymerase (a 5’-exo-minus, N-terminal deletion of Taq DNA polymerase) and a small amount of a proofreading DNA polymerase. This unique blending helps to improve the fidelity, yield and processivity of the resultant PCR process. The Klen-Taq is also highly robust, showing high tolerance of varying concentrations of Mg2+; it is highly thermostable and has four times the fidelity compared to Taq DNA polymerase. The ExcelTaq™ Klen-Taq DNA Polymerase is ideal for DNA amplifications 0.5-5 kb in length on genomic DNA, and up to 10 kb on less complex templates.

N-(Amino-PEG1)-N-bis(PEG2-propargyl) HCl salt is a crosslinker consisting of an amino group with two propargyl groups. The amino group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl groups can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

N-(Amino-PEG1)-N-bis(PEG2-propargyl) HCl salt is a crosslinker consisting of an amino group with two propargyl groups. The amino group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl groups can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.