Introduction

This product provides a reliable solution for DNA isolation from yeast samples. Total DNA can be purified from yeast (<5x 10⁷) without phenol or chloroform. The whole extraction can be finished within 60 minutes. Purified DNA can be directly used for PCR, Southern blot, ect.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from yeast cultures
Applications	PCR, southern blot and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Yeast culture
Sample amount	Bacterial culture: 1-1.5 ml
Elution volume	≥30μl
Time per run	≤60 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris,pH9.0, 0.5mm EDTA).

Advantages

• Fast – several samples can be extracted in 40 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• Good repeatability – silica technology can obtain ideal results every time
• High recovery – DNA can be recovered at the level of PG
• Sufficient components – lysozyme, protease K, RNase A and glass beads

Kit Contents

Contents	D314702	D314703
Purification Times	50 Preps	250 Preps
Hipure DNA Mini Columns I	50	250
2ml Collection Tubes	50	250
Glass Beads (0.4~0.6mm)	20 g	90 g
Buffer SE	30 ml	150 ml
Lyticase	1.8 ml	5 x 1.8 ml
Buffer ATL	30 ml	150 ml
ReagentDX	500 μl	1500 μl
Buffer DL	30 ml	150 ml
Buffer GW1*	13 ml	66 ml
Buffer GW2*	20 ml	2 x 50 ml
Proteinase K	12 mg	60 mg
Protease Dissolve Buffer	1.8 ml	5 ml
Buffer AE	15 ml	30 ml

Storage and Stability

Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. Buffer ATL may precipitate at low temperature. Dissolve it by 37℃ water bath.

This product provides a reliable solution for DNA isolation from yeast samples. Total DNA can be purified from yeast (

Amine-Tri(3-methoxypropanamide-PEG10-Propargyl) Methane HCl salt is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The amino group (NH2) is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde), etc. Reagent grade, for research use only.

Amine-Tri(3-methoxypropanamide-PEG10-Propargyl) Methane HCl salt is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The amino group (NH2) is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde), etc. Reagent grade, for research use only.

Overview

• Purification and enrichment of intact exosomes from plasma, serum, urine, cell culture media and saliva in less than 30 minutes.
• Versatile sample input ranging from 50 µL to 10 mL
  • Plasma/Serum Exosome Purification Mini Kit (50 µL – 1 mL Plasma/Serum)
  • Plasma/Serum Exosome Purification Midi Kit (1 mL – 4 mL Plasma/Serum)
  • Plasma/Serum Exosome Purification Maxi Kit (4 mL – 10 mL Plasma/Serum)
• Exosome purification is based on Norgen’s proprietary resin separating matrix through exosomes’ surface proteins.
• No precipitation reagents, overnight incubation, protease or coagulant treatments required
• No time-consuming ultracentrifugation, filtration or special syringes required
• Purify intact exosomes with a size ranging from 40-200 nm depending on sample input type
• Purified exosomes are compatible with functional studies.
• Purified exosomes are free from any RNA-binding proteins
• Purified exosomes are compatible with NanoSight® or Electron Microscopy for assessing the approximate exosome size range and concentration.
• Exosomal RNA can be extracted from the purified exosomes using Norgen’s Exosomal RNA Purification technology or any other RNA extraction method.

The Plasma/Serum Exosome Purification Kits provide a fast, reliable and convenient method to purify and enrich for intact exosomes from different plasma/serum sample volumes ranging from 50 µL to 10 mL. These kits also allow for the purification of intact extracellular vesicles (EVs) from different plasma/serum sample volumes, and these EVs are ready for any downstream application. The purification is based on Norgen’s proprietary resin.

These kits provide a clear advantage over other available methods since they do not require any special instrumentation, ultracentrifugation, precipitation reagents or any protease treatments. More importantly, the purified exosomes will not be contaminated with any other RNA-binding proteins that may contaminate your exosomal RNA, which is essential if studying exosomal transcripts.

NanoSight® Analysis

Exosomes enriched with Norgen’s Plasma/Serum Exosome Purification Kits can be analyzed using NanoSight® for assessing the approximate exosome size range and concentration

Exosomal RNA Analysis

To purify exosomes and isolate exosomal RNA, choose the Plasma/serum Exosome Purification and RNA isolation kits. The protocol is divided into 2 parts and an aliquot of purified exosomes can be taken for applications like NTA/TEM etc. before processing them for RNA isolation. Or you can use the Exosomal RNA Isolation Kit if you’ve already purified exosomes using a Norgen kit or another method. . Exosomal RNA isolation is based on Norgen’s proprietary resin without the need for phenol extractions or carrier RNA. This RNA is ideal for gene expression analysis using RT-qPCR, microarray, or NGS and for biomarker discovery.

Details

Supporting Data

Figure 1 / 4

Previous

Next

Click for expanded view

Kit Specifications
Plasma/Serum Input (Cat. 57400)	50 μL – 1 mL
Plasma/Serum Input (Cat. 57500)	1 mL – 4 mL
Plasma/Serum Input (Cat. 57600)	4 mL – 10 mL
Size of Exosomes Purified	40 nm – 150 nm
Elution Volume	Variable depending on the plasma/serum input volume
Time to Complete 10 Purifications	15 – 30 minutes

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

Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

Component	Cat. 57400 (50 preps)	Cat. 57500 (25 preps)	Cat. 57600 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	8 mL	2 x 8 mL
ExoR Buffer	12 mL	12 mL	12 mL
Mini Filter Spin Columns inserted into 2 mL tubes	50	25	15
Product Insert	1	1	1