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NGS FFPE DNA Library Prep Kit (illumina platform)

NGS FFPE DNA Library Prep Kit (illumina platform)

NGS FFPE DNA Library Prep Kit (illumina platform)

Product catalog summary
Product Overview
The NGS FFPE DNA Library Prep Kit is designed for constructing high-quality libraries from 10 ng to 400 ng of DNA isolated from formalin-fixed, paraffin-embedded (FFPE) samples. The kit addresses DNA damage caused by fixation, optimizing the repair process for better library quality.
Index Types
  • Non-index: Libraries without index (Cat.# 30035S and 30035L).
  • Index: Unique barcode sequence for library identification, allowing multiplexing of up to 48 samples (Cat.# 30037S and 30037L).
  • Unique Dual Index: Allows multiplexing of up to 96 samples with a 4-Base Difference Index System to minimize sequencing errors (Cat.# 30039S and 30039L).
Features
  • Fast processing time: Total 1.5 hours, with 10 minutes hands-on time.
  • Easy procedure with ready-to-use master mix and fewer reaction components.
  • Reduced magnetic bead requirement by over 50%.
  • High library conversion efficiency with low input DNA requirement.
Kit Components
The kit includes buffers, enzymes, primers, sodium chloride, and PCR mix, with specific volumes varying by catalog number. It requires storage at -20°C for up to 12 months.
Required Equipment (Not Included)
  • Magnetic particle concentrator
  • PCR thermal cycler
  • 96-well PCR plate
  • 80% ethanol
  • Sodium Chloride Solution (1.67 M)
  • Magnetic Beads
Protocol Summary
  1. End Polishing: Mix DNA with FF1 Buffer and Enzyme, incubate, and proceed to adaptor addition.
  2. Adaptor Addition: Add FF2 Buffer and Enzyme, incubate, and add Sodium Chloride.
  3. Pre-PCR Purification: Use magnetic beads for purification, wash with ethanol, and elute library.
  4. PCR: Amplify library with primers and PCR mix, following specified thermal cycling conditions.
  5. Post-PCR Purification: Repeat purification process to obtain the final library.
Quality Control and Usage
The kit components have passed stringent quality tests. It is intended for research and in vitro use only, with specific licensing restrictions on commercial use.
Contact Information
BioDynami, 601 Genome Way, Huntsville, Alabama 35806, USA. For support, contact [email protected].
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Catalog excerpts

NGS FFPE DNA Library Prep Kit (illumina platform)-1

The NGS FFPE DNA Library Prep Kit (illumina platform) was developed for the construction of high quality 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. Three index types are available for the kit: Non-index (Cat.# 30035S and 30035L): Libraries do not have index. Index (Cat.# 30037S and 30037L): Each of our index primers contains a unique barcode sequence with 6 bases that can be used to identify libraries. Library multiplexing up to 48 samples is possible. Unique dual index (Cat.# 30039S and 30039L): Library multiplexing up to 96 samples is possible with unique dual indexes. We have developed a 4-Base Difference Index System. The system allows us to make indexes that have at least 4 bases different from each other in the 8 bases index length. Our unique dual indexing primers remove sequencing errors such as index hopping, index cross-contamination, mis-assignment of reads, amplification errors, and demultiplexing errors. The primer set includes 96 pre-mixed unique pairs of i5 and i7 index primers in a 96-well plate. • Easy procedure ■ Less reaction components • Less magnetic beads required: Reduced more than 50% • Guaranteed quality: Higher library conversion efficiency

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NGS FFPE DNA Library Prep Kit (illumina platform)-2

*Index primers **Unique dual index primers Storage Condition • Store kit at -20°C, stable up to 12 months. J Reagent & Equipment Needed (not provided in this kit) • Magnetic particle concentrator • 80% ethanol (prepare before use) • Sodium Chloride Solution (1.67 M)*** • Magnetic Beads (BioDynami Cat.# 40051) or equivalent ***Sometimes the tube of Sodium Chloride (1.67 M) may crack during dry ice shipping. Customers need to prepare Sodium Chloride (1.67 M) in this case. Library and Index Information Non-Index (Cat.# 30035S and 30035L) Primer sequences: Sequence of the final library with index...

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NGS FFPE DNA Library Prep Kit (illumina platform)-3

For Cat.# 30037S, primers will be shipped in 8-stripe PCR tubes with index labels at both ends as shown below. For Cat.# 30037L, primers will be shipped in 96-well plates. Below is the index layout. Unique Dual Index (Cat.# 30039S and 30039L) Sequence of the final library with index locations: Note: i5 index: NNNNNNN (in yellow) is the index sequence, 5' to 3' direction. I7 index: NNNNNNN (in red) is the index sequence, 5' to 3' direction. List of indexes can be downloaded from: The 96 unique dual index primers have been aliquot in the 96-well plate as shown on left.

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NGS FFPE DNA Library Prep Kit (illumina platform)-4

BioDynamiStep 1: End polishing 1) Add the following to one well of a 96-well PCR plate: 3) Incubate at 20°C for 15 min, 70°C for 5 min. Proceed immediately to step 2. Step 2: Adaptor addition 1) Add the following to Step 1 reaction mixture. Slow pipetting of the viscous FF2 Buffer is needed for precise aliquot. 4) Add Sodium Chloride (1.67 M) 18 ul to the reaction mixture. Proceed immediately to step 3. Step 3: Pre-PCR purification 1) Resuspend Magnetic Beads and transfer 40 ul to the above reaction mixture, mix by pipetting and incubate for 3 min. 2) Load the plate on a magnet, incubate for...

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NGS FFPE DNA Library Prep Kit (illumina platform)-5

Quality Control Kit components passed stringent functional quality test. Product Use Limitation This product is developed and sold for research purposes and in vitro use only. Please refer to BioDynami.com for Material Safety Data Sheet of the product. Limited Label License The product is developed and sold exclusively for research purposes and in vitro use only. The product or its any individual component has not been tested for use in diagnostics or drug development, and is not suitable for administration to human or animal. The purchaser of this product is granted a limited, non-transferable...

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