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.

Introduction

This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples (including serum and plasma). The kit is based on super paramagnetic particles purification technology. Purified DNA/RNA is ready for downstream applications such as Real Time PCR, biochip analysis, NGS and other related experiments.

Details

Specifications

Features	Specifications
Main Functions	Extract Pathogen RNA/DNA from 0.5ml plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution for mNGS application, remove host background nucleic acid.
Applications	RT-PCR，Real Time PCR, biochip analysis, NGS
Products	Pathogen DNA / RNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	low/cell-free samples such as plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution
Sample amount	0.5 ml
Adaptive instrument	Nucleic acid extractor, pipetting workstation

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. After adding magnetic particles and binding solution, DNA/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 DNA/RNA was eluted by Buffer NFW.

Kit Contents

Contents	R667200B	R6672-02B
Purification Times	24 Preps	96 Preps
2ml Bead Tubes	24	96
Proteinase K	12 mg	50 mg
Protease Dissolve Buffer	1.8 ml	3 ml
Buffer SDS (20%)	1.8 ml	8 ml
MagBind Particles	0.6 ml	2.5 ml
Buffer MLB	15 ml	60 ml
Buffer MW1*	13 ml	44 ml
Buffer MW2*	6 ml	50 ml
Buffer AVE	5 ml	30 ml

Storage and Stability

MagBind Particles and Proteinase K Solution 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 18 months under these conditions.

This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples (including serum and plasma). The kit is based on super paramagnetic particles purification technology. Purified DNA/RNA is ready for downstream applications such as Real Time PCR, biochip analysis, NGS and other related experiments.

Overview

• Quantified standard to be used as a positive control or PCR quantification standard
• Vigorously quantified using multiple methods

Neisseria gonorrhoeae is a Gram-negative coccus of the Neisseria genus. N. gonorrhoeae is usually seen in pairs infecting human cells. It has a circular DNA genome of approximately 1Mbp encoding over 2000 genes. N. gonorrhoeae is the etiological agent of the sexually transmitted infection (STI) gonorrhoea, which globally causes an estimated 60 million new cases of gonococcal disease annually. It is second only to Chlamydia trachomatis as the most reported notifiable sexually transmitted disease. Infections with N. gonorrhoeae are primarily restricted to the mucus membranes of the endocervix, urethra, rectum, and pharynx. In females, gonorrhoea is a major cause of pelvic inflammatory disease and may lead to tubal infertility, ectopic pregnancy, and chronic pelvic pain, whereas in males it primarily causes urethritis. Importantly, these infections may often be asymptomatic, thereby contributing to further transmission and maintenance of the disease within populations.

Norgen’s Neisseria gonorrhoeae Quantified Bacterial DNA Standard is prepared from pelleted bacteria grown on culture plates using Norgen’s sample preparation technology. The purified DNA is quantified vigorously using multiple methods including spectrophotometry, gel densitometry and real-time PCR. It is intended to be used as a positive control or PCR quantification standard for Neisseria gonorrhoeae.

Details

Supporting Data

Figure 1 / 1

Previous

Next

Click for expanded view

Volume Provided – 250 µL
DNA Quantity –  2 x 10⁴ copies per µL

Storage Conditions
Upon receipt, store Norgen’s Neisseria gonorrhoeae Quantified Bacterial DNA Standard at -20^oC or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20^oC or lower.