[DM2300] ExcelBand™ 100 bp+3K DNA Ladder, 500μl

The NGS Low Input DNA Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality NGS libraries with low input DNA amount from 1 ng to 400 ng. The kit allows scientist to study samples with limited DNA such as tumor samples, patient samples, and other specially collected samples (FACS sorting etc.). The kit has high library conversion efficiency with as little as 1 ng DNA input. The fast and simple 1.5-hour protocol makes libraries with even coverage and low GC-bias based our unique chemistry for DNA end-polishing and ligation.

The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library preparation, and is compatible with DNA fragments generated from both enzymatic approach (for example, BioDynami DNA fragmentation enzymes, Cat.# 40061 and Cat.# 40062) and mechanical approaches such as sonication and nebulization etc.

NGS Low Input DNA Library Prep Kit Workflow

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

Non-index (Cat.# 30022): Libraries do not have index.

Index (Cat.# 30024): Each of our index primers contains a unique barcode sequence with 6 bases that can be used to identify the low input DNA libraries. Library multiplexing up to 48 samples is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30025): Library multiplexing up to 96 low input DNA libraries is possible with unique dual indexes with our Four-Base Difference Index System. The system allows us to make indexes for the libraries that have at least 4 bases different from each other in the 8-base barcode length. Our unique dual index primers can reduce sequencing errors such as de-multiplexing errors, amplification errors, mis-assignment of reads, index cross-contamination, and also index hopping. The kit  includes 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34024).

Kit advantages:

• • • Fast protocol
      • The hands-on time is only 10 minutes
      • The total protocol time is around 1.5 hours
    • Simple procedure
      • Ready-to-use master mix makes it simple for reaction setup
      • Less reaction components
    • Less magnetic beads required for cleanup steps: Save the cost more than 50%
    • Low input DNA amount: Starts from 1 ng of DNA

Comparison of library conversion efficiency under the same NGS library preparation condition. Input DNA amounts are 1 ng and 10 ng, respectively. DNA was mechanical sheared with Covaris before library prep. BioDynami kit: NGS Low Input DNA Library Prep Kit (Cat. #30022).

Comparison of library yield under the same NGS library prep condition. Input DNA amounts are 1 ng, 10 ng, and 100 ng. DNA was mechanical sheared with Covaris before library prep. BioDynami kit (Cat. #30022). PCR cycle numbers were indicated.

Overview

• Isolate total DNA and RNA from all microorganisms found in water, including bacteria, fungi and algae
• RNA and DNA are both column purified simultaneously using the same column
• Elution contains concentrated DNA and RNA without the need for further precipitation
• Complete RNA (including microRNA) without phenol
• Isolated RNA and DNA are of high quality and integrity for all downstream applications
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a convenient and rapid method for the isolation of microorganisms from water samples. The kits allows for the rapid isolation and purification of total RNA and DNA simultaneously from the microorganisms found in different types of water samples. The total RNA and DNA (including genomic DNA) are isolated from all the microorganisms found in the water, including bacteria, fungi and algae without the use of any inhibitory organic substances The kit purifies genomic DNA, and all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The purified RNA and DNA are highly concentrated, and can be used directly in a number of downstream applications including PCR, qPCR, RT-PCR, qRT-PCR, Northern blotting, Southern blotting and sequencing reactions.

Water RNA/DNA Purification Kit (Spin Column + Filters Kits)

These kits are available with 0.45 μm or 0.22 μm filter formats for capturing microorganisms present in the water.

Water RNA/DNA Purification Kit (Spin Column)

This kit is sold without filters columns.

Details

Supporting Data

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Kit Specifications
Minimum Water Input	10 mL
Maximum Water Input	100 mL
Maximum Filter Column Loading Volume	20 mL
Maximum Spin Column Loading Volume	650 µL
Elution Volume	100 µL
Time to Complete 10 Purifications	45 minutes

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years from the date of shipment.

Component	Cat. 26400 (25 preps)	Cat. 26450 (25 preps)	Cat. 26480 (50 preps)
Lysis Buffer E	15 mL	15 mL	2 x 15 mL
Wash Solution A	18 mL	18 mL	38 mL
Enzyme Incubation Buffer B	6 mL	6 mL	6 mL
Elution Buffer H	6 mL	6 mL	6 mL
Mini Spin Columns	25	25	50
Filter Columns (0.22 μL)	25	–	–
Filter Columns (0.45 μL)	–	25	–
Bead Tubes	25	25	50
Collection Tubes	25	25	50
Elution Tubes (1.7 mL)	25	25	50
Product Insert	1	1	1

Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The 16S V1-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.

The 16S V1-V2 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V1-V2 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

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Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V1-V2 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.

All kit components should remain stable for at least 1 year when stored at the specified storage conditions.

Step	Component	Cat. 70100 (24 preps)	Cat. 70110, 70120, 70130, 70140 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-V2 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL