The ChIP-Seq Library Prep Kit (illumina and MGI Platform) was developed for the construction of high quality ChIP-Seq libraries using 5 ng to 400 ng of ChIP DNA as input. The kit is compatible with ChIP DNA fragments generated from both enzymatic methods and physical methods (sonication, nebulization etc.).
ChIP-Seq Library Prep Kit Workflow
ChIP-Seq is the combination of chromatin immunoprecipitation (ChIP) with next generation sequencing. It is a powerful tool for the analysis of global transcription factors and other proteins in diseases and biological pathways, and characterization of histone modifications in a genome-wide level at single-base resolution. ChIP-Seq delivers whole genome level of functional profiling of global transcription factors, and provides better understanding of epigenetic modifications.
Three index types are available for the ChIP-Seq Library Prep Kit of the illumina platform:
Non-index (Cat.# 30032): Libraries do not have index.
Index (Cat.# 30034): Each index primer contains a unique 6-base index sequence can be used for identification. 48 samples can be pooled together. Index information can be downloaded here.
Unique dual index (Cat.# 30036): The ChIP-Seq library multiplexing for 96 samples is possible. Our unique 4-Base Difference Index System have 8 bases index length and at least 4 bases are different from each other for better library identification. Our unique dual indexing primers remove sequencing errors such as index hopping, index contamination, mis-assignment, and other errors. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34034).
Kit advantages:
Comparison of library conversion efficiency under the same condition. Input DNA amounts are 5 ng and 30 ng. BioDynami ChIP-Seq Library Prep Kit (Cat.# 30034) was used.
Comparison of aligned reads, aligned rate and duplication rate. Input DNA amounts are 5 ng and 30 ng. BioDynami ChIP-Seq Library Prep Kit (Cat.# 30034) was used.
Data comparison: Input DNA amounts are 5 ng and 30 ng. BioDynami ChIP-Seq Library Prep Kit (Cat.# 30034) was used. Sequencing peak regions are shown.
The ChIP-Seq Library Prep Kit (illumina and MGI Platform) was developed for the construction of high quality ChIP-Seq libraries using 5 ng to 400 ng of ChIP DNA as input. The kit is compatible with ChIP DNA fragments generated from both enzymatic methods and physical methods (sonication, nebulization etc.).
The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.
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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.
There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.
Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.
The magnetic beads technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.
Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.
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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.
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Norgen’s 2X One-Step RT-PCR Master Mix is a ready-to-use solution that contains components required for RT-PCR amplification of RNA templates. The mix includes M-MuLV reverse transcriptase, Taq DNA polymerase, dNTPs, reaction buffer, MgCl2, KCl, and a PCR enhancer/stabilizer. The user needs only to add the template, the primer set and water to the Master Mix to set up the RT-PCR reaction. This convenient 2X One-Step RT-PCR Master Mix reduces the time required to set up PCR reactions and reduces the possibility of contamination, particularly when preparing large numbers of reactions. The optimized master mix allows for robust amplification of RNA templates with high yields of PCR products.
Taq DNA Polymerase is a highly thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. The source of Taq included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1. M-MuLV Reverse Transcriptase is an RNA-directed DNA polymerase that can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. The source of the Reverse Transcriptase included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned reverse transcriptase gene from M-MuLV.
Norgen’s 2X One-Step RT-PCR Master Mix is available in 3 convenient sizes:
Cat # 28113 – 100 reactions (sufficient for 100 reactions x 20 µL reaction volume)
Cat # 28114 – 200 reactions (sufficient for 200 reactions x 20 µL reaction volume)
Cat # 28115 – 500 reactions (sufficient for 500 reactions x 20 µL reaction volume)
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Reagents supplied – Cat # 28115
Storage Conditions and Product Stability
2X One-Step RT-PCR Master Mix should be stored at -20°C. For everyday use an aliquot can be stored at 4°C for up to 3 months. Repeated freeze-thaw cycles are not recommended. When stored at the proper temperature this reagent is stable for at least 1 year.
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