The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses 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 (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
Detail
The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses 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 (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
PCR-Free NGS DNA Library Prep Kit Workflow
PCR amplification is a standard step for library preparation of Next-Generation Sequencing. The PCR is used to amplify fully ligated DNA fragments and to add index information to the libraries. The indexing is for the pooling of the library samples in an effort to reduce the sequencing cost.
However, PCR introduces uneven amplification of DNA in some DNA regions with extreme GC-contents and secondary structures. This bias can cause very low sequencing coverage in such regions. DNA sequencing in these regions is still a huge challenge.
PCR-free library prep can reduce library bias and minimize sequencing gaps. The sequencing data from PCR-free library samples have even genomic coverage with few gaps and better depth in GC-rich regions. Our PCR-free NGS kit offers optimal coverage in the regions that are traditionally difficult, such as high-GC content regions, low-GC content regions, and repetitive sequence regions.
Two index types are available for the kit:
Non-index: Libraries do not have index.
Index: Each library contains one i5 index and one i7 index. Library multiplexing up to 96 samples is possible. List of indexes can be downloaded Here.
Kit advantages:
Total time: 1 hr
Hands-on time: 5 min
Easy procedure: Ready-to-use master mix & Less reaction components
Input DNA amount from 100 ng to 1 ug
Other Products
D3129 HiPure Insect DNA Kit
Product Info
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Product Info
Introduction
HiPure Insect DNA Kits provides a simple and rapid solution for total DNA extraction of insect tissue samples. This kit is based on silica gel column purification technology without toxic phenol chloroform extraction and time-consuming alcohol precipitation. The whole extraction process only takes 30 minutes. HiPure Insect DNA Kit can process tissue samples less than 10mg at a time. Hipure Insect DNA 96 kit can process 96 insect tissue samples at a high throughput. The obtained DNA can be directly used in PCR, Southern blot, viral DNA detection and other experiments.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from <10 mg insect tissue
Applications
PCR, southern bolt and virus detection, etc
Purification method
Midi spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Insect tissue samples
Sample amount
<10 mg
Elution volume
≥15μl
Time per run
≤30 minutes
Liquid carrying volume per column
800μl
Binding yield of column
100μg
Principles
This product is based on silica column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
High purity DNA – can be used in sensitive downstream applications such as multiplex and quantitative pcr
Fast – several samples can be extracted in less than 30 minutes
Good repeatability – suitable for extracting high-yield DNA from insect tissue samples
Safe – without phenol chloroform extraction and alcohol precipitation
Kit Contents
Contents
D312902
D312903
Purification Times
50
250
HiPure DNA Mini Columns I
50
250
2ml Collection Tubes
50
250
Buffer ITL
30 ml
120 ml
Buffer IL*
30 ml
120 ml
Buffer GW1*
22 ml
110 ml
Buffer GW2*
20 ml
2 x 50 ml
Proteinase K
24 mg
120 mg
Protease Dissolve Buffer
1.8 ml
15 ml
Buffer AE
15 ml
60 ml
Storage and Stability
Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2–8°C, but in thiscase buffers should be redissolved before use. Make sure that all buffers areat room temperature when used.
Document
HiPure Insect DNA Kits provides a simple and rapid solution for total DNA extraction of insect tissue samples. This kit is based on silica gel column purification technology without toxic phenol chloroform extraction and time-consuming alcohol precipitation. The whole extraction process only takes 30 minutes. HiPure Insect DNA Kit can process tissue samples less than 10mg at a time. Hipure Insect DNA 96 kit can process 96 insect tissue samples at a high throughput. The obtained DNA can be directly used in PCR, Southern blot, viral DNA detection and other experiments.
Plasma/Serum Circulating and Exosomal RNA Purification Kits (Slurry Format)
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Product Info
Overview
Isolate all sizes of circulating and exosomal RNA, including microRNA
Versatile plasma and serum input volumes
Concentrate circulating and exosomal RNA into small elution volumes
Isolate inhibitor-free circulating and exosomal RNA
Bind and elute all RNA irrespective of size or GC content, without bias
Available in Spin Column or 96-well plate formats
Process 96-well plates using vacuum or centrifugation
Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Compatible with Streck Cell-Free RNA BCT® Tubes
Purification is based on Norgen’s proprietary resin matrix
These kits are able to isolate all sizes of circulating and exosomal RNA, including microRNA, without the use of phenol or chloroform. The slurry format provides an advantage over other available kits in that it does not require extension tubes for the purification of free-circulating and exosomal RNA from large sample volumes. RNA can be isolated from either fresh or frozen samples using this kit. This kit is suitable for the isolation of RNA from serum or plasma prepared from blood collected only on either EDTA or citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications including RT-PCR. The purified plasma/serum free-circulating and exosomal RNA is eluted in an elution solution that is compatible with PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
Free-circulating plasma and serum RNA can serve as both tumor- and fetal-specific markers for cancer detection and prenatal diagnosis. As well, free-circulating RNAs have the potential to provide biomarkers for other disease states. Free-circulating RNA in plasma or serum are usually present as short fragments of less than 1000nt, and free-circulating miRNA (21nt) can also be found in plasma and serum.
Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry)
Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating RNA and exosomal RNA from plasma and serum samples ranging from 0.25 mL to 5 mL.
Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Mini Slurry)
Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating and exosomal RNA from plasma and serum samples ranging from 0.25 mL to 2 mL.
Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification 96-Well Kit (Slurry Format) provides a high throughput method for isolating circulating and exosomal RNA from plasma and serum samples using either a vacuum manifold or centrifugation.
Plasma/Serum Circulating and Exosomal RNA Purification Maxi Kit (Maxi Slurry)
Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Maxi Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating RNA and exosomal RNA from various amounts of plasma/serum ranging from 2 mL to 5 mL.
Cat.29500 Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
