The NGS DNA Library Prep Kit was developed for construction of high quality libraries for next generation sequencing (illumina platform). The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods and mechanical methods. Three index types are available for the kit: Non-index (Cat.# 30009S and 30009L): Libraries do not have index. Index (Cat.# 30021S and 30021L): 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.# 30023S and 30023L): 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 • Input DNA amount: From 100 ng to 1 ug

*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 (2 M)*** • Magnetic Beads (BioDynami Cat.# 40051) or equivalent ***Sometimes the tube of Sodium Chloride (2 M) may crack during dry ice shipping. Customers need to prepare Sodium Chloride (2 M) in this case. Library and Index Information Non-Index (Cat.# 30009S and 30009L) Primer sequences: Sequence of the final library with index location:...

For Cat.# 30021S, primers will be shipped in 8-stripe PCR tubes with index labels at both ends as shown below. For Cat.# 30021L, primers will be shipped in 96-well plates. Below is the index layout. Unique Dual Index (Cat.# 30023S and 30023L) 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.

BioDynamiProtocolStep 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 DL2 Buffer is needed for precise aliquot. 4) Add Sodium Chloride (2 M) 40 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...

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...