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

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.).

Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Midi Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine.

This product is for research use only.

Video

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 1-5ml plasma, serum, body fluids using vacuum protocol
Applications	qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (vacuum)
Sample type	Serum, plasma and other cell-free fluid samples
Sample amount	1-5ml
Elution volume	≥50μl
Time per run	≤60 minutes
Liquid carrying volume per column	4ml
Binding yield of column	1mg

Principle

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.

Advantages

• High yield – most optimal process, free DNA (>50bp) can be obtained to the maximum extent
• High concentration – low elution volume, ensuring high nucleic acid concentration
• High purity – low alcohol binding method, completely removing inhibitor and protein pollution
• High recovery – DNA can be recovered at thelevel of PG by silica gel column purification

Kit Contents

Contents	D318201C	D318202C
Purification Times	10	50
Buffer ACL	50 ml	250 ml
Buffer ACB*	60 ml	300 ml
Buffer DCW1*	4.4 ml	22 ml
Buffer DCW2*	5 ml	10 ml
Proteinase K	110 ml	540 mg
Protease Dissolve Buffer	10 ml	30 ml
Carrier RNA	110 μg	110 μg
Nuclease Free Water	10 ml	20 ml
HiPure CFDNA Mini Columns	10	50
2 ml Collection Tubes	20	100
Extender Tube	10	50
Vac-Connector	10	50

Storage and Stability

Proteinase K, Carrier RNA 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 dry 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 this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

Experiment Data

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,

Name of Product

Aspergillus Antigen
(Galactomannan) – Rapid Test

Catalog Number

ASP-025

Short Info

This Rapid Test is for the detection of specific Aspergillus Galactomannan Antigen in Serum or Bronchoalveolar Lavage (BAL).

This product is manufactured by GaDia Diagnostics in Switzerland and distributed in Germany exclusively by Milenia Biotec.

Method/Platform

Immunochromatographic Rapid Test

Range/Assay Sensivity

Test Principle

The principle of the test is colloidal gold immunochromatography. If the sample is positive, the antigens in the sample react with the red-colored nanoparticles and form a complex (Antigen – anti-Aspergillus monoclonal antibodies – gold nanoparticles), which was previously pre-dried on the conjugate pad. The mixture then moves upward on the membrane by capillary action. As the sample flows through the test membrane, the binding conjugate complexes migrate. The anti-Aspergillus antibodies present on the membrane (Test Line) capture the colored conjugate complex and a red line will appear.

Brief Instructions

Sample Preparation and Result Interpretation

Storage

2-8° C

Components

Aspergillus Galactomannan Detection Test, Positive Control, Negative Control, Sample Treatment Solution

25 tests
This Rapid Test is for the detection of specific Aspergillus Galactomannan Antigen in Serum or Bronchoalveolar Lavage (BAL).

This product is manufactured by GaDia Diagnostics in Switzerland and distributed in Germany exclusively by Milenia Biotec.