endo-BCN-PEG4-Biotin

Overview

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Isolate all sizes of circulating DNA from plasma and serum samples
• Isolate viral and bacterial DNA and RNA
• Versatile plasma and serum input volumes (10 µL – 200 µL)
• No phenol extractions
• Bind and elute all RNA irrespective of size or GC content, without bias
• Concentrate circulating RNA, exosomal RNA and cell-free circulating DNA into a flexible elution volume ranging from 10 µL to 25 µL
• Isolate inhibitor-free nucleic acids
• Purify high-quality RNA and DNA in 30 minutes
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Norgen’s Plasma/Serum RNA/DNA Purification Mini Kit provides a fast, reliable, reproducible and simple procedure for the sequential isolation of circulating RNA, exosomal RNA and Cell-Free Circulating DNA (cfc) from the same Plasma/Serum sample. It can sequentially isolated RNA and DNA from small plasma/serum input ranging from 10 µL to 200 µL. The kit is designed to isolate all sizes of circulating RNA, including microRNA, all sizes of exosomal RNA as well as all sizes of cfc-DNA from fresh or frozen plasma or serum samples. Norgen’s Plasma/Serum RNA Purification Kit provides a clear advantage over other available kits in that it does not require Phenol/Chloroform or any Protease treatments for the isolation of plasma/serum RNA or DNA . RNA and DNA can be eluted into a flexible elution volume ranging from 10 µL to 25 µL. Purified RNA can be used in a number of sensitive downstream applications including reverse transcription qPCR, reverse transcription PCR, NGS, Northern blotting, RNase protection and primer extension, and expression array assays. Purified DNA can be used in a number of sensitive downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis.

Background

Typical yields of free-circulating, exosomal RNA and cfc-DNA vary depending on the input sample, as the amount of RNA and/or DNA present in plasma and serum will depend upon the health status of the individual.  Normally, the RNA/DNA yield from plasma or serum is highly variable (range from 1 to 100 ng/mL). Variability is also observed between samples collected from the same donor at different times during the day. This kit is suitable for the isolation of RNA/DNA from 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.

Details

Supporting Data

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Kit Specifications
Minimum Plasma/Serum Input Volume	10 μL
Maximum Plasma/Serum Input Volume	200 μL
Size of RNA Purified	All sizes including small RNA (<200 nt)
Size of DNA Purified	≥ 50 bp
Minimum Elution Volume	10 μL
Maximum Elution Volume	25 μL
Time to Complete 10 Purifications	15-20 minutes
Average Yield	Variable depending on specimen

 Note: Do not exceed the recommended sample input volume of 200 µL.

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.

Component	Cat. 55200 (50 preps)
Lysis Buffer A	30 mL
Wash Solution A	38 mL
Solution WN	18 mL
Elution Solution A	6 mL
Elution Buffer B	8 mL
Micro Spin Columns	50
Micro-Elute RNA Spin Columns	50
Collection Tube	100
Elution Tubes (1.7 mL)	100
Product Insert	1

Propargyl-PEG4-amine has a propargyl group and an amine group. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls, etc. The propargyl group can form triazole linkage with azide-bearing biomolecules. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG4-amine has a propargyl group and an amine group. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls, etc. The propargyl group can form triazole linkage with azide-bearing biomolecules. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The 16S V1-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V3 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.

The 16S V1-V3 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V1-V3 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

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Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V1-V3 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.

All kit components should remain stable for at least 1 year when stored at the specified storage conditions.

Step	Component	Cat. 70200 (24 preps)	Cat. 70210, 70220, 70230, 70240 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-V3 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL