The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Detail
Introduction
The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Details
Specifications
Features
Specifications
Main Functions
Isolation miRNA from 0.3-0.5ml serum or plasma using magnetic particles
Applications
RT-PCR, cDNA synthesis, second generation sequencing
Purification method
Polydisperse magnetic beads
Purification technology
Magnetic beads technology
Process method
Manual or automatic
Adaptive instrument
Nucleic acid extractor, pipetting workstation
Sample type
Serum, plasma, acellular samples
Sample amount
300μl
Principle
This product is based on the purification method of high binding magnetic particles. The sample material is denatured in Lysis Buffer. The protein is then precipitated by Protein Precipitation Solution and pelleted by centrifugation. After adding magnetic particles and binding solution, RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption.The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally RNA was eluted by Elution Buffer.
Advantages
Safe – no phenol chloroform extraction and no use of Trizol reagent
Fast – several samples can be extracted in 60 minutes by column method
Kit Contents
Contents
R662801
R662802
R662803
Purification Times
48 Preps
96 Preps
5 x 96 Preps
MagPure Particles N
1.7 ml
3.5 ml
17 ml
Buffer CFL
6 ml
12 ml
60 ml
Buffer CPL
1.8 ml
3.5 ml
20 ml
Buffer MGW1*
30 ml
60 ml
250 ml
Buffer MW2*
12 ml
25 ml
100 ml
RNase Free Water
10 ml
20 ml
60 ml
Storage and Stability
MagPure Particle N should be stored at 2–8°C upon arrival. However, short-term storage (up to 8 weeks) at room temperature (15–25°C) does not affect their 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.
Other Products
Sulfo DBCO-Maleimide
Product Info
Document
Product Info
Sulfo DBCO-Maleimide a water-soluble sulfhydryl reactive containing a maleimide group and a DBCO moiety. Maleimide group specifically and efficiently reacts with thiols to form stable thioether bonds. The low mass weight will add minimal spacer to modified molecules and will enable simple and efficient incorporation of DBCO moiety onto cysteine-containing peptides or other thiol-containing biomolecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
Sulfo DBCO-Maleimide a water-soluble sulfhydryl reactive containing a maleimide group and a DBCO moiety. Maleimide group specifically and efficiently reacts with thiols to form stable thioether bonds. The low mass weight will add minimal spacer to modified molecules and will enable simple and efficient incorporation of DBCO moiety onto cysteine-containing peptides or other thiol-containing biomolecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
AAV Purification from any input – cell fraction or media fraction
High AAV recovery, up to 90%
No specialized equipment needed
Purification from a variety of AAV serotypes (including AAV6 and AAV9)
Yields highly active AAV for in vivo and in vitro experiments
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
Recombinant adeno-associated virus (AAV) vectors are highly promising tools for both in vitro and in vivo gene transfer. Norgen’s AAV Purification Kits provide fast and simple procedures for concentrating and purifying AAV vectors from cell lysate and cell culture media. Purification is based on precipitation onto Norgen Biotek’s proprietary resin. Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. AAV vector purified in this manner is highly active for use in in vitro and in vivo transduction experiments.
AAV Purification Kit
Norgen’s AAV Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant (SN) or a total of 500 mL of supernatant input). Approximately 1 mL of cell pellet can be purified per prep, up to a maximum of 15 mL of cell pellet in total for the entire kit. Up to 33X sample concentration.
AAV Purification Mini Kit
Each spin column is able to concentrate and purify AAV from 0.5-8 mL of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (200 µL). Preparation time for 4 samples is 1.5 hours, with 45 minutes of hands-on time.
AAV Purification Midi Kit
Each spin column is able to concentrate and purify AAV from 8 mL up to 45 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1 mL). The kit may be used to purify up to 8 x 25 mL or 4 x 45 mL of samples using the included columns. Preparation time for 4 samples is approximately 2 to 2.5 hours, with 1.5 hours of hands on time.
AAV Purification Maxi Kit (Slurry Format)
Each spin column is able to concentrate and purify AAV from 45 mL to 90 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 200X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1-10 mL) using the optional concentration step. The kit may be used to purify up to 1 x 900 mL samples or 10 x 45-90 mL samples using the included columns. Preparation time for 1 x 900 mL sample is approximately 2.5 to 3.5 hours, with an optional concentration step requiring an additional 30 min.
At least 5 x 109 AAV particles as determined by qPCR
AAV Vector Serotype
Any
Average Recovery
> 80%
Input Type
Cells, media, or mixed
Input Volume
0.5 mL – 8 mL
Minimum Elution Volume
200 µL
Time to Complete Purifications
1 – 2 hours
Storage Conditions and Product Stability DNAse I and RNAse A should be stored at -20°C upon arrival. Elution Buffer O should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 1 year after the date of shipment.
Component
Cat. 66100 (15 preps)
Cat. 63200 (20 preps)
Cat. 63300 (4-8 preps)
Cat. 63250 (1-10 preps)
Lysis Buffer S
5.5 mL
5.5 mL
5.5 mL
20 mL
DNAse I
–
2 x 25 uL
2 x 25 uL
210 μL
RNAse A
–
60 μL
60 μL
240 μL
HL-SAN Nuclease
102 μL
–
–
–
Binding Buffer A
20 mL
4 mL
4 mL
2 x 8 mL
Purification Solution C
60 mL
–
–
–
Purification Solution D
130 mL
–
–
–
Wash Solution C
2 x 130 mL
60 mL
60 mL
3 x 60 mL
Slurry E
12.5 mL
–
–
2 x 14.5 mL
Elution Buffer O
66 mL
8.5 mL
8.5 mL
66 mL
Protein Neutralizer
4 mL
4 mL
4 mL
4 mL
Spin Columns
–
20
–
–
Mini Spin Columns
–
20
–
–
Midi Spin Columns (grey contents) with Collection Tubes
–
–
8
10
Midi Spin Columns (white contents) with Collection Tubes
–
–
8
–
Maxi Spin Columns (grey contents) with Collection Tubes
–
–
–
10
Maxi Spin Columns (white contents) with Collection Tubes
t-Boc-aminooxy-PEG5-propargyl is a click crosslinker. Propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. T-Boc-aminooxy can be converted to free aminooxy under mild acidic conditions and then can react with an aldehyde or ketone. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
t-Boc-aminooxy-PEG5-propargyl is a click crosslinker. Propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. T-Boc-aminooxy can be converted to free aminooxy under mild acidic conditions and then can react with an aldehyde or ketone. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
