Propargyl-PEG4-t-butyl ester

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

• Fast and easy high throughput processing using either a vacuum manifold or centrifugation
• Process all soil types including clay, loam, sandy soils and high humic content soils such as peat, compost and manure
• Remove organic substances using the OSR Solution
• Remove all humic acid from DNA samples using the Humic Acid Removal plate
• Isolate high quality total DNA from all soil types – ready for any downstream PCR, qPCR
• ​Excellent DNA for metagenomic studies
• No phenol or chloroform extractions

This kit provides a fast, reliable and simple procedure for high throughput isolation of DNA from all types of soil samples including common soil samples and difficult soil samples with high humic acid content such as compost and manure. A combination of chemical and physical homogenization effectively lyses all microorganisms in the soil sample, and the kit removes all traces of humic acid using the provided Organic Substance Removal (OSR) Solution and Humic Acid Removal plate (HAR). Total genomic DNA can be isolated and purified from all the various microorganisms found in soil, such as bacteria, fungi and algae. The purified DNA is of the highest quality and is fully compatible with downstream PCR and qPCR  applications and more for any metagenomic study, as all humic acid substances and PCR inhibitors are removed during the isolation.

Details

Supporting Data

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Kit Specifications
Binding Capacity Per Well	50 μg
Maximum Loading Volume Per Well	500 μL
Size of DNA Purified	All sizes
Maximum Amount of Starting Material	250 mg
Time to Complete 96 Purifications	50 minutes

Storage Conditions and Product Stability
All 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. 26560 (192 preps)
Lysis Buffer G	2 x 100 mL
Lysis Additive A	25 mL
Binding Buffer I	25 mL
OSR Solution	12 mL
Wash Solution A	2 x 38 mL
Elution Buffer B	30 mL
Bead Tubes	200
HAR Plate	2
96-Well Filter Plate	2
Adhesive Tape	4
96-Well Collection Plate	4
96-Well Elution Plate	2
Product Insert	1