Ethanol Assay Kit (Liquid Ready)

The NGS FFPE DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality FFPE DNA libraries using 10 ng to 400 ng of input DNA isolated from formalin-fixed, paraffin-embedded (FFPE) samples. The DNA damage caused by fixation makes it difficult to construct high quality libraries. Our kit has been optimized to repair damaged DNA in the reactions. Multiplexing of the NGS FFPE DNA Library is possible.

NGS FFPE DNA Library Prep Kit Workflow

FFPE samples are a great resource for biomedical research. However, the methods for fixation and condition of storage significantly damage the DNA in the samples.  Thus, the extraction of high quality DNA from FFPE samples is often a challenge.  Low yield and low quality of FFPE DNA usually are common because of the limited tissue material and the DNA degradation.

As a result, it is usually difficult to construct high quality NGS libraries from low amount and low quality of FFPE DNA. In order to address this issue, we have developed the NGS FFPE DNA Library Prep Kit to make high quality libraries from the low input of FFPE DNA samples.

Three index types are available for the NGS FFPE DNA Library Prep Kit of the illumina platform:

Non-index (Cat.# 30035): Libraries do not have index.

Index (Cat.# 30037): Each of our index primers contains a unique barcode DNA (6 bases long) that can be used to identify individual library. Multiplexing of libraries is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30039): FFPE DNA library multiplexing is possible with 96 samples based on the unique dual indexing system. Our unique Four–Base Difference Index System allows us to make indexes that have at least 4 bases different from each other in the 8-base index sequence. Our unique dual indexing primers can effectively remove NGS errors including index hopping, de-multiplexing errors, index cross-contamination, mis-assignments etc. The unique dual index primer set includes 96 pre-mixed unique pairs of i5 and i7 index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34037).

Kit advantages:

• Fast and simple protocol
  • Making libraries in just 1.5 hours
  • 10 minutes of hands-on time
• Easy procedure
  • Ready-to-use master mix to reduce the tedious mixing
  • Less reaction components to simplify the reaction setup
• Less magnetic beads needed for the purification steps: saving more than 50% of the expensive beads
• Low input FFPE DNA: From 10 ng to 400 ng

Comparison of library conversion efficiency under the same condition. Input DNA amount is 25 ng.

Comparison of library yield under the same condition. Input DNA amount is 25 ng.

The NGS FFPE DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality FFPE DNA libraries using 10 ng to 400 ng of input DNA isolated from formalin-fixed, paraffin-embedded (FFPE) samples. The DNA damage caused by fixation makes it difficult to construct high quality libraries. Our kit has been optimized to repair damaged DNA in the reactions. Multiplexing of the NGS FFPE DNA Library is possible.

Overview

• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias
• Versatile sample input ranges
• Isolate all sizes of free-circulating RNA, including microRNA​
• Bind and elute all RNA irrespective of size or GC content, without bias
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents nor overnight incubation required
• Concentrate isolated exosomal RNA and free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
• 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 sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits since they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Urine Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 250 µL to 1 mL.

Urine Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 2 mL to 10 mL.

Urine Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 11 mL to 30 mL.

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	2 mL
Maximum Urine Input	10 mL
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50 – 100 µL
Time to Complete 10 Purifications	35 – 40 minutes
Average Yields*	Variable depending on the specimen

*Please check page 5 of the product insert for the average yields and the common RNA quantification methods.

Storage Conditions and Product Stability
All buffers 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.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Component	Cat. 59200 (50 preps)	Cat. 59300 (25 preps)	Cat. 59400 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	2 x 20 mL	2 x 20 mL	2 x 20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	2 x 18 mL	18 mL	18 mL
Elution Solution A	2 x 6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	100	50	30
Collection Tubes	100	50	30
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1