This product provides a reliable solution for DNA isolation from yeast samples. Total DNA can be purified from yeast (
Detail
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.
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads(tRNA Purification) to overcome the hurdle.
The Magnetic Beads(tRNA Purification) solves the tRNA and short DNA/RNA purification problem. The beads with our proprietary technology purify tRNA effectively by removing impurities and unwanted components such as dNTPs, detergents, salts, proteins, and other contaminants. The magnetic bead reagents can be used for oligo (>70 nt) applications.
Workflow without large RNA/DNA contamination
In the case of samples contaminated with RNA/DNA such as rRNA and DNA, our magnetic beads can effectively remove RNA/DNA that are 180 nt and larger. Purified tRNA are ideal for applications requiring high quality, as the fragments are free of impurities and contaminants.
Workflow with large RNA/DNA contamination
Comparison of tRNA and 70 nt oligos recovery. BioDynami beads (tRNA purification) successfully recovered DNA fragments both yeast tRNA and 70 nt oligos.
Recovery of tRNA and 70 nt oligos with BioDynami magnetic Beads (tRNA Purification). Yeast tRNA and 70 nt oligos were used as input. Input and recovered oligos were quantified with ssDNA Quantification kit (BioDynami Cat. # 40043) and RNA Quantification kit (BioDynami Cat. # 40044).
Depletion of larger RNAs. Left panel: depletion of 28S rRNA and 18S rRNA. Right panel: depletion of RNA of 180 nt and larger.
Features
Removal of unwanted components and other impurities
Purification of tRNA and oligos (>70 nt)
tRNA
RNA fragments 70 nt or longer
DNA/RNA hybrid fragments 70 nt or longer
Oligo and chimeric oligo 70 nt or longer
dsDNA fragments 70 bp or longer
ssDNA fragments 70 nt or longer
Removal of larger RNA/DNA contamination:
18S rRNA
28S rRNA
RNA/DNA> 180 nt
Document
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads (tRNA Purification) to overcome the hurdle.
Isolate all sizes of circulating and exosomal RNA, including microRNA
Versatile plasma/serum input ranges
No phenol extractions
No carrier RNA
Bind and elute all RNA irrespective of size or GC content, without bias
Concentrate circulating RNA and exosomal RNA into a flexible elution volume
High quality, purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Compatible with Streck Cell-Free RNA BCT® Tubes
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples 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. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
Background
Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as plasma or serum.
Plasma/Serum RNA Purification Mini Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 µL to 200 µL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 µL to 25 µL.
Plasma/Serum RNA Purification Midi Kit
This utilizes a two column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 µL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
Plasma/Serum RNA Purification Maxi Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
*This kit is suitable for the isolation of RNA 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.
**Please check page 7 for Average Plasma/Serum Yields and Common RNA Quantification Methods.
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
