Uvod

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 minuta. 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.

pojedinosti

Tehnički podaci

Načelo

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, itd.

Prednosti

• Brzo – može se izdvojiti nekoliko uzoraka 20 minutes by column method
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Sef – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG
• Visok prinos – carrier RNA contained in the product maximize the recovery of trace nucleic acid

Sadržaj kompleta

Skladištenje i stabilnost

Carrier RNA should be stored at 2–8°C upon arrival. Međutim, kratkotrajno skladištenje (do 12 tjedni) na sobnoj temperaturi (15–25°C) ne utječe na njihovu izvedbu. Preostale komponente kita mogu se čuvati na sobnoj temperaturi (15–25°C) i stabilni su najmanje 18 months under theseconditions