DBCO-C5-acid

Overview

• Sequentially isolate and purify total RNA and DNA from a single sample
• Two column system: one for DNA and one for RNA
• The RNA column is for the purification of total RNA including microRNA
• No need to split the lysate, or to use phenol or precipitation methods
• Rapid and efficient spin column procedure – it takes only 30 minutes
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a rapid method for the isolation and purification of total RNA and DNA sequentially from a single sample of cultured animal cells and tissues, blood, bacteria, yeast, or fungi. The lysate is passed over two columns: 1) a DNA column and 2) an RNA column. Total RNA of all sizes is purified, including microRNA. Both DNA and RNA are of the highest purity and yield.

These kits are ideal for researchers who are interested in studying the genome and transcriptome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/DNA Purification Kits are especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as they eliminate the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and DNA are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications

RNA/DNA Purification Kit (Spin Column)

Maximum column binding capacity of 50 μg for RNA and 20 μg for DNA.

RNA/DNA Purification Micro Kit (Micro)

The purified RNA and DNA fractions can be eluted in as little as 20 μL. Ideal for cell number inputs of 500,000 and as little as 5 cells. Maximum column binding capacity of 35 μg for RNA and 10 μg for DNA.

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	50 μg for RNA 20 μg for DNA
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Maximum Amount of Starting Material: Animal Cells Animal Tissues Blood Bacteria Yeast Fungi Plant Tissues	5 x 106 cells 25 mg (for most tissues)** 200 μL 1 x 109 cells 1 x 108 cells 50 mg 50 mg
Time to Complete 10 Purifications	30 minutes
Average Yield*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg)	10-15 μg RNA 2-4 μg DNA 30-35 μg RNA 4-6 μg DNA

*Average Yield will vary depending upon a number of factors including species, growth conditions used, and development stage.

**Tissue inputs of up to 40 mg may be used, however for inputs greater than the recommended 25 mg, cross-contamination of the RNA and DNA fractions is possible.

Storage Conditions and Product Stability
Store Proteinase K at -20°C upon arrival. All other 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. 48700 (50 preps)	Cat. 50300 (50 preps)
Buffer SKP	40 mL	40 mL
Wash Solution A	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL
Elution Solution A	6 mL	6 mL
Elution Buffer F	15 mL	6 mL
RNase-Free Water	40 mL	40 mL
Proteinase K	2 x 12 mg	2 x 12 mg
gDNA Purification Columns	50	–
gDNA Purification Micro Columns	–	50
RNA Purification Columns	50	–
RNA Purification Micro Columns	–	50
Collection Tubes	100	100
Elution Tubes (1.7 mL)	100	100
Product Insert	1	1

Description

The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5′ end of RNA to form m7Gppp5’N-RNA (Cap-0 RNA). The 2′-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2′ -OH of the first nucleotide at the 5′ end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.

Features

• 2’-O-Methyltransferase included for Cap-1 RNA
• High capping efficiency
• High stability
• RNase inhibitor is included to enhance the stability of capping reaction

Application

• Generation of 5’Cap-0 (m7Gppp) and Cap-1 (m7GpppNm-) RNA by enzymatic reaction
• mRNA synthesis for in vitro translation
• Gene expression studies
• mRNA vaccine development and therapeutics

Storage

-20°C for 24 months

The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5′ end of RNA to form m7Gppp5’N-RNA (Cap-0 RNA). The 2′-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2′ -OH of the first nucleotide at the 5′ end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.