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DNA Library Prep Kit for IIlumina Sequencing

To prepare DNA samples for Illumina Sequencing
Catalog #: K1475
$425.00

Product Details

Cat # +Size K1475-12
Size 12 Rxns
Kit Summary DNA library is prepared for Illumina sequencing by end repair, adenylation, ligation with adapter sequences and amplification
Detection Method DNA library is prepared for Illumina Sequencing by end repair, adenylation, ligation with adapter sequences and PCR amplification
Applications Library preparation for Whole Genome Sequencing, ChIP-Seq, Amplicon Sequencing and cDNA Sequencing
Features & Benefits • High quality Illumina compatible libraries
• Minimal hands on time and short 2 hour library preparation workflow
• Wide range of input amounts (2-500 ng) and sample types, including amplicons and fragmented DNA
Kit Components • 10X End Repair Buffer
• End Repair Enzyme Mix
• Ligation Enhancer
• Ligation Buffer
• Ligation Enzyme Mix
• Index Adapters
• Magnetic Purification Beads
• PCR Primer Mix
• 2X PCR MasterMix
• Nuclease-free water
Storage Conditions -20ºC
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

BioVision’s DNA Library Prep Kit for IIlumina Sequencing is used to efficiently convert (2 ng-500 ng) of DNA fragments into fully compatible libraries for Illumina Next Generation Sequencing (NGS) platforms. The sample preparation step involves the preparation of a DNA library by fragmentation, end repair, adenylation, ligation with adapter sequences and amplification. The DNA library is then loaded onto the chips for amplification and sequencing. The presence of adapter sequences allows the hybridization of the libraries to the sequencing chips and provides a universal priming site for sequencing primers. BioVision’s DNA Library Prep Kit contains index adapters needed for running multiple samples on a single Illumina flow cell as well as the magnetic beads for purification, which is required for the reaction cleanup after ligation and PCR steps. The entire procedure can be completed within 2 hours.


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The sample preparation for NGS platforms involves preparation of a DNA library by fragmentation, end repair, adenylation, ligation with adapter sequences and amplification. Does this kit include any steps for adenylation as shown in the figure.1 in the datasheet?
Yes. The End Repair Reaction step in the kit protocol includes the adenylation step to prime the ends for adapter ligation.
How should I fragment the DNA before using the kit?
The DNA samples can be fragmented by mechanical shearing using Focused ultrasonicator or enzymatically using enzymes such as DNA Fragmentase.
How many adapters will be provided with the kit?
The kit will contain 12 vials of adapters. Each vial will contain one adapter.
The following sentence in the datasheet is not clear. "If multiple samples are being prepared at the same time, use a different Index Adapter for each sample". Please explain.
If 12 samples are prepared for sequencing together, please use a different Index Adapter vial for each sample.
Can this kit be used for cDNA samples?
Yes. The kit can be used for cDNA samples.
Please let us know what does the PCR Primer Mix code for?
The barcodes (index sequences) are included in the adapters. The PCR Primer Mix simply amplifies the Adapter-ligated fragments that contain the Illumina P5 and P7 sequences from the adapters that are required for binding to the Illumina Sequencing flow cell.
In the magnetic beads based purification with dual size selection, we add beads to the DNA and discard the beads containing some DNA for the first time. The DNA that did not bind to the beads is present in the supernatant. Then Magnetic beads are added to the supernatant again. Will the DNA that did not bind to the beads previously bind to the beads during the second time?
The ratio of magnetic beads to DNA samples, specifically the concentration of Polyethylene Glycol (PEG) determines the size of DNA fragments that will bind to the beads and be retained. When the magnetic beads are added for the first time at the 0.8X ratio (40 μl beads:50.5 μl Adapter Ligation Reaction Mixture Volume), they will retain fragments larger than 200-300 bp. The unbound DNA will contain the smaller fragments resulting from primer dimers and adapter dimers. If more magnetic beads are added to the unbound DNA, it may bind to the beads due to increasing concentrations of PEG. But it is undesirable for sequencing.