Cell Culture Flask 5 Layers

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 V3-V4 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V3-V4 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 V3-V4 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 V3-V4 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

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 V3-V4 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. 70400 (24 preps)	Cat. 70410, 70420, 70430, 70440 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V3-V4 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

Overview

• Fast and easy high throughput processing using either a vacuum manifold or centrifugation
• Process all soil types including clay, loam, sandy soils and high humic content soils such as peat, compost and manure
• Remove organic substances using the OSR Solution
• Remove all humic acid from DNA samples using the Humic Acid Removal plate
• Isolate high quality total DNA from all soil types – ready for any downstream PCR, qPCR
• ​Excellent DNA for metagenomic studies
• No phenol or chloroform extractions

This kit provides a fast, reliable and simple procedure for high throughput isolation of DNA from all types of soil samples including common soil samples and difficult soil samples with high humic acid content such as compost and manure. A combination of chemical and physical homogenization effectively lyses all microorganisms in the soil sample, and the kit removes all traces of humic acid using the provided Organic Substance Removal (OSR) Solution and Humic Acid Removal plate (HAR). Total genomic DNA can be isolated and purified from all the various microorganisms found in soil, such as bacteria, fungi and algae. The purified DNA is of the highest quality and is fully compatible with downstream PCR and qPCR  applications and more for any metagenomic study, as all humic acid substances and PCR inhibitors are removed during the isolation.

Details

Supporting Data

Figure 1 / 2

Previous

Next

Click for expanded view

Kit Specifications
Binding Capacity Per Well	50 μg
Maximum Loading Volume Per Well	500 μL
Size of DNA Purified	All sizes
Maximum Amount of Starting Material	250 mg
Time to Complete 96 Purifications	50 minutes

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 26560 (192 preps)
Lysis Buffer G	2 x 100 mL
Lysis Additive A	25 mL
Binding Buffer I	25 mL
OSR Solution	12 mL
Wash Solution A	2 x 38 mL
Elution Buffer B	30 mL
Bead Tubes	200
HAR Plate	2
96-Well Filter Plate	2
Adhesive Tape	4
96-Well Collection Plate	4
96-Well Elution Plate	2
Product Insert	1

Product advantages

1. The instantaneous centrifugal function rises to the maximum speed for about 6 seconds, allowing for rapid detachment of hanging droplets from the wall;

2. Faster acceleration and deceleration rate, required to complete the experiment in a shorter time;

3. Door cover protection, overspeed and imbalance detection system, capable of real-time monitoring of  the centrifugal process, ensuring the safe operation    of the  instrument; When the operation ends, an error occurs, or an imbalance occurs, the sound signal prompts, and the operation stops at the same time. The LCD displays the result code

Host Parameters

Product model	Mini-3
Input power supply	DC24V/2.75A
Input power	55W
Motor/Drive Method	DC24V brushless DC variable frequency motor
Display method	High brightness LCD screen
Effective centrifugal time	1-99min.1-59sec
speed	300~3000±15rpm
Speed step increase	10rpm
Maximum capacity	2 *96 well PCR plates/ELISA plates
MAX. RCF	608 xg
Misoperation/alarm failure	Sound prompt+display code
weight	3.9Kg
Noise	60dB (A)
Fastest acceleration time	<6s
Fastest deceleration time	<5s
Permissible ambient temperature/relative temperature	+5-40°C/80%

The micro porous plate centrifuge adopts international advanced design concepts and manufacturing technology, with a smooth and beautiful appearance, compact and stable structure, adhering to the advantages of high universality and easy operation. It also has the functional characteristics of “smooth start” and “smooth braking”; This micro porous plate centrifuge is very suitable for 96 or 384 well and small capacity micro porous plates, as well as various standard PCR micro porous plates with or without skirts .