DBCO-PEG6-amine TFA salt

Overview

• Excellent separation and purification of cytoplasmic and nuclear RNA
• Convenient and fast spin column format
• High quality and yield of RNA
• Isolate full diversity of RNA (including microRNA) without phenol
• Purified RNA is ready for any application including RT-PCR, qRT-PCR, RNA-Seq, arrays and more
• Cytoplasmic RNA is free of DNA and ready for direct use in RT-PCR/qRT-PCR
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol.   Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations.   Ten samples can be processed in approximately 45 minutes.  This kit is also available in a 100 prep size.    

Background

In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA. 

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	Up to 50 µg RNA
Maximum Column Loading Volume	650 µL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Time to Complete 10 Purifications	45 minutes
RNA Yield HeLa (1 x 106) – Cytoplasmic RNA HeLa (1 x 106) – Nuclear RNA	15 µg ≤ 3.5 µg

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.

Component	Cat. 21000 (50 preps)	Cat. 37400 (100 preps)
Lysis Buffer J	20 mL	2 x 20 mL
Buffer SK	40 mL	2 x 40 mL
Wash Solution A	38 mL	2 x 38 mL
Elution Buffer E	6 mL	2 x 6 mL
Mini Spin Columns	50	100
Collection Tubes	50	100
Elution Tubes (1.7 mL)	50	100
Product Insert	1	1

Description

The ExcelTaq™ Hot Start II DNA Polymerase is a mixture of an aptamer-based inhibitor and a recombinant thermo-stable Taq DNA polymerase designed for preventing or minimizing non-specific DNA amplification in PCR reaction. The inactivation of polymerase is achieved by a reversible binding of the aptamer to the polymerase at temperatures below 45°C. The aptamer inhibitor releases polymerase during normal PCR cycling. The aptamer-based inhibition omits the time-consuming initial activation step required by chemically modified or antibody-based hot start polymerases.

The high specificity and sensitivity of ExcelTaq™ Hot Start II DNA Polymerase allows sensitive detection from limited amount of DNA templates, such as 1 pg of cDNA or 1 fg of plasmid DNA. With a high DNA synthesis rate and high thermo-stability, the ExcelTaq™ Hot Start II DNA Polymerase allows reactions to be set up at room temperature and is suitable for common and specialized PCR applications

Features

• Aptamer-based hot start PCR
• Reversible enzyme inactivation
• Omits extra enzyme activation step
• Convenient for room temperature PCR set-up
• High yield and specificity of target amplicons
• Wide range of amplicon length (up to 10 kb)
• High sensitivity (as low as 1 fg of plasmid)

Applications 

• High specificity PCR
• Generation of PCR products for TA cloning
• Routine PCR, multiplex PCR, colony PCR, and RT-PCR

Storage

-20°C for 24 months 

The ExcelTaq™ Hot Start II DNA Polymerase is a mixture of an aptamer-based inhibitor and a recombinant thermo-stable Taq DNA polymerase designed for preventing or minimizing non-specific DNA amplification in PCR reaction. The inactivation of polymerase is achieved by a reversible binding of the aptamer to the polymerase at temperatures below 45°C. The aptamer inhibitor releases polymerase during normal PCR cycling. The aptamer-based inhibition omits the time-consuming initial activation step required by chemically modified or antibody-based hot start polymerases.

The high specificity and sensitivity of ExcelTaq™ Hot Start II DNA Polymerase allows sensitive detection from limited amount of DNA templates, such as 1 pg of cDNA or 1 fg of plasmid DNA. With a high DNA synthesis rate and high thermo-stability, the ExcelTaq™ Hot Start II DNA Polymerase allows reactions to be set up at room temperature and is suitable for common and specialized PCR applications

Overview

• Purify sequencing extension products from dye terminators, primers and other contaminants
• Also purify DNA from different enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations.
• High recovery
• Fast and efficient spin column format
• Also available in 96 well format for high throughput

This kit provides a rapid spin column procedure for the purification and clean-up of sequencing and various other enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations. The kit is used to remove reaction contaminants including dye terminators, salts, enzymes, excess primers and primer dimers. Contaminants are undesirable as they can interfere with many downstream applications including sequencing, RFLP, restriction enzyme digestions and ligation. Purification is based on spin-column chromatography without the use of phenol, chloroform or alcohol precipitation. The kit provides a high quality product with up to 90% recovery.

The kit is also available in a 96-well format for high-throughput sequencing reaction clean-up.  Purification with the 96-well plate can be performed using either a vacuum manifold or centrifugation.

Details

Supporting Data

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Kit Specifications – Spin Column
Column Binding Capacity	13 μg
Maximum Column Loading Volume	650 μL
Maximum Amount of Starting Material	25 μg
Time to Complete 10 Purifications	10 minutes
Minimum Elution Volume	30 μL

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 34500 (50 preps)	Cat. 34400 (2 x 96-well plate)
Binding Buffer C	30 mL	3 x 30 mL
Wash Solution A	12 mL	2 x 20 mL
Elution Buffer B	8 mL	2 x 15 mL
Spin Columns	50	–
Collection Tubes	50	–
Elution Tubes (1.7 mL)	50	–
96-Well Plate	–	2
Adhesive Tape	–	4
96-Well Collection Plate	–	2
96-Well Elution Plate	–	2
Product Insert	1	1