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NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms)

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The NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality cell-free DNA (cfDNA) libraries using 1 ng to 50 ng of cell-free DNA as input. The kit has a simple work flow and a fast procedure. Multiplexing of the cell free DNA library is possible based on the index type.

Detail

The NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality cell-free DNA (cfDNA) libraries using 1 ng to 50 ng of cell-free DNA as input. The kit has a simple work flow and a fast procedure. Multiplexing of the cell free DNA library is possible based on the index type.

NGS Cell Free DNA Library Prep Kit (illumina)

NGS Cell Free DNA Library Prep Kit Workflow

The main source of cell-free DNA (cfDNA) is derived from apoptotic hematopoietic cells in blood and found in the plasma. The length of the cfDNA is about 150-200 bp in length. Circulating tumor DNA (ctDNA) derived from malignant tumors is a part of cfDNA. Both cfDNA and ctDNA can be used as a noninvasive biomarker since it offers a better approach in comparison to invasive tissue biopsies.

NGS has been used for cfDNA and ctDNA sequencing in the field of liquid biopsy as it provides a whole genome level of molecular profiling. One of the hurdles for cfDNA sequencing is the difficulty of library preparation from the limited amount of cfDNA obtained from plasma. Our cell-free NGS kit makes it easy to get enough libraries from limited input in just 1.5 hours.

Three index types are available for the NGS Cell Free DNA Library Prep Kit of the illumina platform:

Non-index (Cat.# 30029): Libraries do not have index.

Index(Cat.# 30031): Each of our index primers contains a unique 6-base index sequence that can be used for sample identification. Total 48  library multiplexing is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30033): Cell-free DNA library multiplexing up to 96 samples is possible with the unique dual indexes. We have developed a Four-Base Difference Index System. The system have at least 4 bases different from each other in the 8 bases index length. The primers effectively minimize sequencing errors such as mis-assignment, index hopping, index contamination etc. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34031).

Kit advantages:

  • Fast and simple protocol
    • Hands-on time is around 10 minutes
    • The total protocol time is only 1.5 hours
  • Easy procedure based on:
    • Ready-to-use master mix for easy setup of reactions
    • Less reaction components to simplify bench work
  • Less magnetic beads used for cleanup procedures: This can reduce more than 50% of the beads consumption
  • Guaranteed high library conversion efficiency
  • Low input cell-free DNA: From 1 ng to 50 ng

NGS Cell Free DNA Library Prep Kit

Comparison of library conversion efficiency with different samples under the same condition.

NGS Cell Free DNA Library Prep Kit

Comparison of library yield with different samples under the same condition.

Other Products

Fungi/Yeast Genomic DNA Isolation Kits

Overview

  • Rapid spin column purification of genomic DNA from viable yeast cells, fungal spores or mycelium, and bacteria including Gram-positive
  • Bead tubes (provided) allow for effective mechanical homogenization
  • Purified DNA is of high quality and integrity and compatible with any sensitive downstream applications such as PCR, qPCR, RFLP and more

These kits provide fast and reliable procedures for the purification of genomic DNA from viable yeast cells, fungal spores or mycelium, and bacteria including Gram-positive. Genomic DNA is efficiently extracted from the cells by a combination of heat treatment, detergents and Bead Tubes (provided). An optional lyticase treatment allows for improved DNA yields with certain fungal and yeast species. Recovered genomic DNA is of excellent yield and purity for any downstream application including PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), Amplified Fragment Length Polymorphism (AFLP), sequencing, SNP analysis and more.

Fungi/Yeast Genomic DNA Isolation Kit (Spin Column)

This kit provides rapid spin column purification of genomic DNA from viable yeast cells, fungal spores or mycelium, and bacteria including Gram-positive. Preparation time for a single sample is less then 30 minutes, and each kit contains sufficient materials for 50 preparations.

Fungi/Yeast Genomic DNA Isolation 96-Well Kit (HT)

Norgen’s Fungi/Yeast Genomic DNA Isolation 96-Well Kit provides a fast, reliable and simple procedure for high throughput isolation of DNA from viable yeast cells, fungal spores or mycelium and Gram-positive bacteria. The purification could be performed on either a vacuum manifold or using centrifugation. Complete 96 purifications in 40 minutes.

Details

Supporting Data

Figure 1 / 6

Previous

Figure 1. DNA Isolation from Different Fungi Species and Yeast
Figure 1. DNA Isolation from Different Fungi Species and Yeast
Figure 2. DNA can be Isolated from Spores and Mycelium
Figure 3. Identification of Fungal and Yeast Species in a Real-time PCR (SYBR Green)
Figure 4. High Yields of DNA Isolated from Saccharomyces cerevisiae overnight culture
Figure 5. High quality of yeast DNA measured by NanoDrop
Figure 6. Purified DNA Can be Amplified in a Real-time PCR (SYBER GREEN) Reaction.

Next

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Kit Specifications
Binding Capacity Per Well50 μg
Maximum Loading Volume Per Well500 μL
Size of DNA PurifiedAll sizes
Maximum Amount of Starting Material:
Fungi (Wet weight)
Yeast or Gram-positive bacterial culture
50 mg
0.5 mL – 1 mL
Time to Complete 96 Purifications40 minutes

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 2 years in their unopened containers.

ComponentCat. 27300 (50 preps)Cat. 27350 (192 preps)
Lysis Buffer L30 mL2 x 60 mL
Resuspension Solution A20 mL60 mL
Solution BX28 mL2 x 28 mL
Wash Solution A18 mL2 x 38 mL
Elution Buffer B15 mL30 mL
Bead Tubes50200
Spin Columns50
Collection Tubes50
96-Well Plate2
96-Well Collection Plate2
Adhesive Tape4
Elution Tubes (1.7 mL)50
96-Well Elution Plate2
Product Insert11

Cell Culture Flask T25

Cell Culture Flask T25

Cell Culture Flask surface is very smooth based on precise molding technology and it gives clear view when examined with microscope.

Volume: 25mL75mL175mL225mL

PRODUCT FEATURES

  • The product is made of medical grade USP CLASS VI polymer polystyrene
  • The product is made under a 100,00- class dust-free manufacturing site
  • Two kinds of product line up are providing.

For adherent cell culture: Initial adherence and proliferative property of cells via hydrophilic surface treatment.

For suspension cell culture: The surface is resistant to cell adherence, which minimizes damage or loss of cell.

  • Large mouthed design makes easy operation of pipet or cell scraper. The surface of flask is uniform and smooth, hence the clear view can be obtained when microscopic observation.
  • The hydrophobic filter cap can prevent invasion of fungi and bacteria without water absorb.
  • Gamma radiation sterilization
  • Non-Pyrogenic, DNase/Rnase free.

 Magnetic Beads (DNA & RNA Purification)

Magnetic Beads (DNA & RNA Purification)

Magnetic Beads (DNA & RNA Purification), Replacement of SPRI Beads such as AMPure XP & SPRIselect

Solid Phase Reversible Immobilization magnetic beads consist of paramagnetic particles coated with carboxyl groups that reversibly bind DNA. They are used for DNA purification because they are fast, simple and efficient. We have developed our own beads technology that are different from other SPRI beads technologies.

Our Magnetic Beads (DNA & RNA Purification) combines BioDynami’s proprietary chemistries with reversible DNA-binding properties of magnetic beads. The magnetic beads, which are unique from other SPRI beads, are developed for effective nucleic acid purification by removing unwanted components such as salts, dNTPs, enzymes, primers, adapters, and other impurities. The beads are RNase free, can be used for applications of DNA, and even work with more sensitive RNA without any additional cost.

Our magnetic beads are optimized to selectively bind DNA fragments of 100 bp and larger, and RNA fragments of 200 bases and larger, similar to other SPRI beads such as AMPure® XP* and SPRIselect*. Purified DNA and RNA are suitable for downstream applications requiring high quality DNA and RNA, as the purified fragments are free of contaminants and impurities. The beads can be used for NGS library purification, PCR fragment cleanup, molecular cloning, or even nucleic acid concentration.

magnetic beads DNA purification workflow

The beads can also be used for size selection of DNA fragments ranging from 150 bp to 800 bp by changing the bead-to-sample volume ratio and performing single or double-size selection. The beads are an ideal choice for NGS library preparation. They can be easily integrated into the standard workflow of NGS library preparation since the volume ratio is similar for protocols using standard magnetic beads.

Magnetic Beads binding capacity

Features:

  • Effective recovery of DNA and RNA samples
    • DNA fragments greater than 100 base pairs
    • RNA fragments greater than 200 bases
  • Removal of unwanted components and impurities
  • Fragment size selection for specific applications
    • Consistent single or double-size selection
  • Flexibility: compatible with manual and automated processing
  • Cost effective alternative to other beads such as AMPure® XP* and SPRIselect*  with equivalent performance

* AMPure® XP and SPRIselect are trade marks of Beckman Coulter.

SPRI beads recovery rate

Magnetic beads recovery rate. dsDNA and ssDNA of genomic DNA, 1 kb DNA, and 200 bp DNA fragments were used. Total RNA was also tested.

Comparison of elution volume vs yield. 40 ul , 30 ul, and 20 ul of elution volume were used. BioDynami magnetic beads have better recovery rates at low elution volume when compared to other SPRI beads such as AMPure® XP*.

Applications:

  • NGS Library preparation
  • Purification of PCR fragments
  • Purification of DNA and RNA fragments
  • Molecular Cloning
  • Other applications requiring purified DNA and RNA