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

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Kit Specifications
Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Maximum Amount of Starting Material: Fungi (wet weight) Yeast or Bacteria Culture	50 mg 0.5mL – 1mL
Average Yield* Pichia sp. (yeast) Botrytis cinerea ​Fusarium sp. Aspergillis niger Mucor racemosus Cladosporium cladosporioides Penicillium sp. Rhizopus oryzae Alternaria tenuissima	25 μg 32 μg 42 μg 26 μg 15 μg 12 μg 40 μg 7 μg 30 μg
Time to Complete 10 purifications	30 minutes

* Yield will vary depending on the type of sample processed

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. 27300 (50 preps)	Cat. 27350 (192 preps)
Lysis Buffer L	30 mL	2 x 60 mL
Resuspension Solution A	20 mL	60 mL
Solution BX	28 mL	2 x 28 mL
Wash Solution A	18 mL	2 x 38 mL
Elution Buffer B	15 mL	30 mL
Bead Tubes	50	200
Spin Columns	50	–
Collection Tubes	50	–
96-Well Plate	–	2
96-Well Collection Plate	–	2
Adhesive Tape	–	4
Elution Tubes (1.7 mL)	50	–
96-Well Elution Plate	–	2
Product Insert	1	1

Introduction

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.

Details

Specifications

Features	Specifications
Main Functions	Isolation miRNA and other small RNA molecules(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column
Applications	RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal tissues, adherent cells, suspension cells, bacteria, etc
Sample amount	Eukaryotic culture cells: ≤ 10^7, Animal tissue:<100mg, Yeast culture cells:<5 x10^7, Bacteria:<10^9
Elution volume	≥15μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such asguanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein andother impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

This kit combines acid/guanidine (MagZol) extraction technology with glass fiber filter membrane purification, which can improve the extraction effect of complex samples and samples with low RNA content. After the sample is treated with MagZol reagent and chloroform, the supernatant is added with ethanol to provide appropriate binding conditions, then transferred to the purification column and centrifuged. Macromolecular RNA can be efficiently bound to the membrane. Collect the filtrate containing small RNA, add more ethanol to adjust the binding capacity of small RNA, the pollutants can be efficiently washed away by second cleaning. Finally, the purified RNA was eluted by RNase free water.

Advantages

• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – several samples can be extracted in 40 minutes
• High applicability – samples including animals, plants, bacteria, cells, etc.
• High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity

Kit Contents

Contents	R431002	R431003
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	100	2 x 250
2ml Collection Tubes	100	2 x 250
MagZol Reagent	60 ml	270 ml
Buffer RWC	20 ml	80 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

MagZol Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its 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 Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.

Description 

AccuBand™ 100 bp DNA Ladder II is composed of 10 individual DNA fragments, presenting 1k, 900, 800, 700 600, 500, 400, 300, 200 and 100 bp sharp bands respectively. This product contains 1 enhanced band (500 bp) for easy identification of bands. AccuBand™ 100 bp DNA Ladder II is ready-to-use, containing loading buffer with dual color tracking dyes (orange G and Xylene cyanol FF). AccuBand™ 100 bp DNA Ladder II provides a sufficient amount of DNA for clear observation of all DNA bands ranging from 100 bp to 1 kb, either in agarose gel or in polyacrylamide gel electrophoresis. 

Features

• Sharp bands
• Suitable for polyacrylamide gel electrophoresis
• Quick reference— enhanced bands 
• Ready-to-use— premixed with loading dye for direct loading
• Stable— room temperature storage over 6 months

Source 

Phenol extracted PCR products and dsDNA digested with specific restriction enzymes, equilibrated in 10 mM Tris-HCl (pH 8.0) and 10 mM EDTA.

Range 

100 ~ 1,000 bp 

Concentration 

50 µg/ 500 µl 

Recommended loading volume 

5 µl/ well

Storage

Room temperature for 6 months
4°C for 12 months 
-20°C for 36 months