DBCO-PEG2-PFP ester

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

Figure 1 / 4

Previous

Next

Click for expanded view

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

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 V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-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 V2-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 V2-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

Figure 1 / 3

Previous

Next

Click for expanded view

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 V2-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. 70300 (24 preps)	Cat. 70310, 70320, 70330, 70340 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V2-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

Tetra-(amido-PEG10-propargyl) is a branched crosslinker with four terminal propargyl groups. The propargyl groups can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage.

Tetra-(amido-PEG10-propargyl) is a branched crosslinker with four terminal propargyl groups. The propargyl groups can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage.