Overview

• Isolate high quality DNA from a broad variety of phage strains
• High yields of total DNA
• Fast and easy processing using a rapid spin-column format
• No phenol or chloroform extractions or cesium chloride banding required
• High yields of DNA recovered 3-15 µg DNA from 10⁶-10¹⁰ pfu/ mL of enriched phages

This kit provides a rapid spin column method for the purification of total DNA from a broad spectrum of bacteriophages propagated in bacteria grown in liquid cultures. The DNA is isolated without the use of phenol, chloroform or cesium chloride banding procedures. The spin-column based procedure is rapid and can be completed in less than 45 minutes. The kit is highly efficient for processing small volumes of phage supernatant (500 µL – 1 mL) and with the optional DNase and Proteinase K treatments phage DNA yields are maximized while host DNA contamination is minimized. Purified total phage DNA is of the highest integrity, and can be used in a number of downstream applications including PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), sequencing, cloning, Southern Blot and more.

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Details

Supporting Data

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Kit Specifications
Column Binding Capacity	50 µg
Maximum Column Loading Volume	650 µL
Size of DNA Purified	All sizes
Maximum Amount of Starting Material	1 x 1010 pfu/mL enriched phages
Average Yield*	3-15 µg DNA from 106-1010 pfu/mL of enriched phages
Time to Complete 10 Purifications	45 minutes

* Average yields will vary depending upon a number conditions used and developmental stage.

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. 46800 (50 preps)	Cat. 46850 (100 preps)
Lysis Buffer B	40 mL	2 x 40 mL
Wash Solution A	38 mL	2 x 38 mL
Elution Buffer B	8 mL	2 x 8 mL
Spin Columns	50	100
Collection Tubes	50	100
Elution Tubes (1.7 mL)	50	100
Product Insert	1	1

Introduction

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

This method is suitable for small quantities of tissue (<100mg) and cells (<5 X10⁶), and large quantities of tissue (up to 1g) and cells (<10⁸), of human, animal, plant, or bacterial origin. The simplicity of the MagZol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. Total RNA prepared in this manner can be used for Northern blot analysis, dot blot hybridization, poly(A) + selection, in vitro translation, RNase protection assay, and molecular cloning. For use in amplification by thermal cycling, treatment of the isolated RNA with RNase-free DNase I is recommended when the two amplimers lie within a single exon.

Details

Specifications

Features	Specifications
Main Functions	Extract RNA from liquid samples by salting out method
Applications	RT-PCR, Northern hybridization, poly (a) enrichment, etc.
Purification technology	Acid phenol guanidine isothiocyanate
Process method	Manual (centrifugation)
Sample type	Various liquid samples
Sample amount	Flexible
Elution volume	Variation with sample size
Time per run	Variation with sample size

Advantages

• Flexible – sample amount can be adjusted according to the demand
• Cost performance -the most economical nucleic acid extraction technology

Storage and Stability

MagZol Reagent should be stored at 2-8°C upon arrival and is stable for at least 24 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance.

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

Overview

• Eight discrete bands, ranging from 100 to 1,000 bases
• Higher intensity band at 500 bases for easy reference
• No need for staining or destaining as loading dye contains ethidium bromide
• Convenient lyophilized format provides better product stability

The Norgen 100 b RNA Ladder is a set of RNA transcripts derived from recombinant DNA templates. This ladder is suitable for precise sizing of small RNA molecules using native or denaturing agarose gels, and can be easily visualized under UV.

Contents

• 2 vials of lyophilized RNA ladder (25 loads each)
• 1x loading buffer: 33.3% formamide, 11.7% formaldehyde, 0.012% bromophenol blue, 0.05mM EDTA, 0.012% ethidium bromide, and 0.7x MOPS solution
• 2x loading buffer

Details

Supporting Data

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Specifications:

• Eight discrete bands, ranging from 100 to 1,000 bases
• Higher intensity band at 500 bases for easy reference

Contents:

• 2 vials of lyophilized RNA ladder (25 loads each)
• 1x loading buffer: 33.3% formamide, 11.7% formaldehyde, 0.012% bromophenol blue, 0.05mM EDTA, 0.012% ethidium bromide, and 0.7x MOPS solution
• 2x loading buffer

Instructions:

To reconstitute the lyophilized RNA ladder, add 250 µL of 1x loading buffer to each 25 loads vial and vortex gently.
Heat at 80°C for 10 minutes. Incubate on ice for 1 min. Load 10 µL on a 1.5-2% gel. For optimal results, use Norgen 2x loading buffer with each RNA sample. There is no need for staining and destaining denaturing gels since Norgen’s loading buffer contains ethidium bromide.

Storage:

Store at -20°C.