Introduction

Hipure viral RNA kit is suitable for purifying viral RNA from samples such as cell-free body fluid or culture medium. The kit is based on silica gel column purification technology. It requires no toxic phenol chloroform extraction and time-consuming alcohol precipitation in the extraction. The whole extraction process takes only 25 minutes. The kit is suitable for extracting viral RNA from 1-140µl cell-free fluid samples such as serum, plasma, cell-free body fluid or culture medium. The product has successfully extracted hepatitis B A/C, hepatitis C RNA, SARS and HIV. The obtained RNA can be directly used for RT-PCR, Northern hybridization and virus detection.

Details

Specifications

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 as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other 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.

The sample is homogenized and lysed in the lysate, and RNA is released into the lysate. The high concentration of guanidine isothiocyanate contained in the lysate denatured and inactivated endogenous or exogenous RNase, while RNA is protected from degradation. The lysate is centrifuged to remove insoluble impurities. After adding ethanol to adjust the binding conditions, it is transferred to the column for filtration. RNA is adsorbed on the membrane of the column, while protein is removed without adsorption. The column is washed with buffer VHB to remove protein and other impurities, washed with buffer RW2 to remove salt. Finally the RNA is eluted by RNase free water. The eluted RNA can be directly used in RT-PCR, Northern blot, poly-A purification, in vitro translation, etc.

Advantages

• Fast – several samples can be extracted in 20 minutes by column method
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG
• High yield – carrier RNA contained in the product maximize the recovery of trace nucleic acid

Kit Contents

Storage and Stability

Carrier RNA 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 at room temperature (15–25°C) and are stable for at least 18 months under theseconditions

Hipure viral RNA kit is suitable for purifying viral RNA from samples such as cell-free body fluid or culture medium. The kit is based on silica gel column purification technology. It requires no toxic phenol chloroform extraction and time-consuming alcohol precipitation in the extraction. The whole extraction process takes only 25 minutes. The kit is suitable for extracting viral RNA from 1-140µl cell-free fluid samples such as serum, plasma, cell-free body fluid or culture medium. The product has successfully extracted hepatitis B A/C, hepatitis C RNA, SARS and HIV. The obtained RNA can be directly used for RT-PCR, Northern hybridization and virus detection.

Overview

• Purification from all sequence cycling by-products
• Purification of all types of enzymatic reactions
• High recovery
• Complete magnetic bead purification
• High integrity product
• Also available in a 96-well format that can be integrated with a robotic automation system
• Clean-up of amplified DNA products from enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations

Norgen’s PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System) provides a rapid, simple and efficient procedure for the purification and clean-up of sequencing and various other enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations. The kit is used to remove reaction contaminants including dye terminators, salts, enzymes, excess primers and primer dimers. Contaminants are undesirable as they can interfere with many downstream applications including sequencing, RFLP, restriction enzyme digestions and ligation. Purification is based on magnetic beads without the use of phenol, chloroform or alcohol precipitation.  The kit provides a high quality product with up to 90% recovery.

Norgen’s PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System) is also available in a 96-well format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.

Details

Supporting Data

Figure 1 / 2

Previous

Next

Click for expanded view

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.

Description 

ExcelTaq™ Klen-Taq DNA Polymerase is a specially blended enzyme mix containing KlenTaq-1 DNA polymerase (a 5’-exo-minus, N-terminal deletion of Taq DNA polymerase) and a small amount of a proofreading DNA polymerase. This unique blending helps to improve the fidelity, yield and processivity of the resultant PCR process. The Klen-Taq is also highly robust, showing high tolerance of varying concentrations of Mg²⁺; it is highly thermostable and has four times the fidelity compared to Taq DNA polymerase. The ExcelTaq™ Klen-Taq DNA Polymerase is ideal for DNA amplifications 0.5-5 kb in length on genomic DNA, and up to 10 kb on less complex templates.

Features

• 5’→3′ DNA polymerase activity
• 3’→5′ exonuclease activity (proofreading)
• 4× fidelity as compared to Taq DNA polymerase
• Thermo-stable: up to 98°C during PCR denaturing step
• Robust PCR performance, resistance to variance in PCR conditions 

Storage

-20°C for 24 months 

ExcelTaq™ Klen-Taq DNA Polymerase is a specially blended enzyme mix containing KlenTaq-1 DNA polymerase (a 5’-exo-minus, N-terminal deletion of Taq DNA polymerase) and a small amount of a proofreading DNA polymerase. This unique blending helps to improve the fidelity, yield and processivity of the resultant PCR process. The Klen-Taq is also highly robust, showing high tolerance of varying concentrations of Mg2+; it is highly thermostable and has four times the fidelity compared to Taq DNA polymerase. The ExcelTaq™ Klen-Taq DNA Polymerase is ideal for DNA amplifications 0.5-5 kb in length on genomic DNA, and up to 10 kb on less complex templates.