ELISA HiPure Viral RNA
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Introduction
Hipure viral RNA kit is suitable for purifying viral RNA from samples such as cell-free body fluid or cµLture 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 cµLture medium. The product has successfµLly 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
Features Specifications
Main Functions Extract viral RNA from 140μl cell-free samples
Applications RT-PCR, Northern hybridization, and various virus detection
Purification method Mini spin column
Purification technology Silica technology
Process method Manual (centrifµgation or vacuum)
Sample type Cell-free body fluid or cµLture medium
Sample amount 140μl
Elution volume ≥15μl
Time per run ≤25 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 as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid throµgh 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 centrifµged 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
Contents R417102 R417103
Purification Times 50 Preps 250 Preps
HiPure Viral Micro Columns 50 250
2mL Collection Tubes
100
500
Buffer VRL 50 mL
200 mL
Carrier RNA 310 µg 3 x 310 µg
Buffer VHB*
13 mL
110 mL
Buffer RW2*
20 mL
2 x 50 mL
Nuclease Free Water
10 mL
30 mL
Storage and Stability
Carrier RNA shoµLd 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
Articles
【JOURNAL OF VIROLOGY】L226Q Mutation on Influenza H7N9 Virus Hemagglutinin Increases Receptor-Binding Avidity and Leads to Biased Antigenicity Evaluation | Journal of Virology
【Engineering】Redesigned Duplex RT-qPCR for the Detection of GI and GII Noroviruses
【Frontiers in Microbiology】Development of a High-Efficiency Immunomagnetic Enrichment Method for Detection of Norovirus via PAMAM Dendrimer/SA-Biotin Mediated Cascade-Amplification
【FOOD CONTROL】Detection of group A rotavirus in oyster tissues by in situ capture RT-qPCR
【Viruses-Basel】Library Preparation Based on Transposase Assisted RNA/DNA Hybrid Co-Tagmentation for Next-Generation Sequencing of Noroviruses
【JOURNAL OF MEDICAL VIROLOGY】GII.17[P17] and GII.8[P8] noroviruses showed different RdRp activities associated with their epidemic characteristics