Acetic Acid Assay Kit (Acetate Kinase Manual Format)

Alkyne-PEG2-iodide is a crosslinker containing a propargyl group and an iodine group. The propargyl group can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry reactions. Iodine (I) is a very good leaving group for nucleophilic substitution reactions. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research use only.

Alkyne-PEG2-iodide is a crosslinker containing a propargyl group and an iodine group. The propargyl group can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry reactions. Iodine (I) is a very good leaving group for nucleophilic substitution reactions. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research use only.

Overview

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Versatile plasma/serum input ranges
• No phenol extractions
• No carrier RNA
• Bind and elute all RNA irrespective of size or GC content, without bias
• Concentrate circulating RNA and exosomal RNA into a flexible elution volume
• High quality, purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Compatible with Streck Cell-Free RNA BCT® Tubes
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.

Background

Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as plasma or serum.

Plasma/Serum RNA Purification Mini Kit

This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 µL to 200 µL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 µL to 25 µL.

Plasma/Serum RNA Purification Midi Kit

This utilizes a two column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 µL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.

Plasma/Serum RNA Purification Maxi Kit

This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.

Isolate RNA after Purifying EVs and Exosomes

For Ultracentrifugation, Exoquick, and Filtration

Cat. #	Name	Elution Volume	Plasma/Serum	Urine	Cell-Culture Media
55000	Plasma/Serum RNA Purification Mini Kit	10 – 25 µL	50 µL – 1 mL	250 µL – 1 mL	5 – 10 mL
35300	Total RNA Purification Micro Kit	20 – 50 µL	1 – 4 mL	2 – 10 mL	10 – 20 mL
17200	Total RNA Purification Kit	50 – 100 µL	4 – 10 mL	11 – 30 mL	20 – 35 mL

Details

Supporting Data

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Kit Specifications
Sample Volume Range	50 to 200 μL
Anti-coagulant (for Plasma)*	EDTA or Citrate
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Minimum Elution Volume	10 μL
Maximum Elution Volume	25 μL
Time to Complete 10 Purifications	15-20 minutes
Average Yield**	Variable depending on specimen

*This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

**Please check page 7 for Average Plasma/Serum Yields and Common RNA Quantification Methods.

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. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Kit Components	Cat. 55000 (50 preps)	Cat. 56100 (20 preps)	Cat. 56200 (10 preps)
Lysis Buffer A	2 x 20 mL	100 mL	1 x 130 mL 1 x 30 mL
Wash Solution A	18 mL	1 x 38 mL 1 x 18 mL	38 mL
Elution Solution A	6 mL	6 mL	6 mL
Elution Buffer F	–	15 mL	15 mL
Micro Spin Columns	50	–	–
Mini Spin Columns	–	20	10
Midi Spin Columns	–	20	–
Maxi Spin Columns	–	–	10
Collection Tubes	50	20	10
Elution Tubes (1.7 mL)	50	20	10
Product Insert	1	1	1

Overview

• No need for staining and destaining of denaturing gels since Norgen’s loading buffer contains ethidium bromide
• Highly stable
• Nine bands from 200 bases to 4000 bases
• Convenient lyophilized format provides better product stability

The Norgen 1 kb RNA Ladder is a set of RNA transcripts derived from recombinant DNA templates. This ladder is suitable for precise sizing of a wide range of RNA molecules using native or denaturing agarose gels, and can be visualized under UV.

Contents: 
• 2 vials of lyophilized RNA ladder (25 loads each)
• 1x loading buffer: 33.3% formamide, 11.7% formaldehyde, 0.012% bromophenol blue, 0.05mM EDTA, 0.012% ethidium bromide, and 0.7x MOPS solution
• 2x loading buffer

Details

Supporting Data

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Specifications:
• Nine discrete bands, ranging from 200 to 4,000 bases
• Higher intensity band at 1,000 bases for easy reference

Contents:
• 2 vials of lyophilized RNA ladder (25 loads each)
• 1x loading buffer: 33.3% formamide, 11.7% formaldehyde, 0.012% bromophenol blue, 0.05mM EDTA, 0.012% ethidium bromide, and 0.7x MOPS solution
• 2x loading buffer

Instructions:
To reconstitute the lyophilized RNA ladder, add 250 µL of 1x loading buffer to each 25 loads vial and vortex gently.
Heat at 80°C for 10 minutes. Incubate on ice for 1 min. Load 10 µL on a 1-2% gel. For optimal results, use Norgen 2x loading buffer with each RNA sample. No need for staining and destaining of denaturing gels since Norgen’s loading buffer contains ethidium bromide.

Store at -20°C.

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