RNA extraction reagent kit using spin-column sorption "RNA Pro Q" Raissol

The kit includes all reagents required for RNA extraction:
1. 3-reagent - vodnyy rastvor fenola i guanidin izotiotsianata dlya bystrogo lizisa kletok;
2. Precipitation buffer - dlya razdeleniya obraztsa na tri fazy: vodnuyu, interfazu i organicheskuyu, pri etom RNA, DNA i belki okazyvayutsya v raznykh fazakh;
3. Binding buffer - dlya povysheniya sorbtsii RNA na filtre kolonki;
4. Wash buffer - dlya promyvki RNA ot cell metabolites and salts;
5. Elution buffer - dlya elyutsii i khraneniya RNA;
6. Spin-kolonki - dlya filtratsii reaktsionnoy smesi i sorbtsii RNA.

Download the short instruction "RNA Pro Q" (pdf)

Download the full instruction "RNA Pro Q" (pdf)

Provedenie protsedury extraction RNA

Lysis
1. V tubes obemom 1,5-2 mL place 10-100 mg obraztsa i add 600 uL 3-reagenta. Pri ispolzovanii zhidkikh obraztsov obem 3-reagenta dolzhen prevyshat obem obraztsa v 10 raz (60 uL obraztsa - 600 uL 3-reagenta).
2. Intensivno peremeshat tubes on a vortex mixer. Incubate at room temperature v techenie 10 minut, periodicheski vstryakhivaya. Sbrosit kapli s pomoshchyu brief centrifugation.
3. Centrifuge tubes pri +4℃ v techenie 10 minut pri 13 tys. ob/min. 4. Akkuratno, ne zadevaya pellet, perenesti supernatant v novye tubes obemom 1,5-2 mL.
Ekstraktsiya i sorbtsiya RNA
1. Add v tubes 100 uL osazhdayushchego bufera, intensivno peremeshivat tubes on a vortex mixer v techenie 15-30 sek.
2. Centrifuge tubes pri +4℃ v techenie 10 minut pri 13 tys. ob/min.
3. Akkuratno, ne zadevaya interfazu, perenesti 250 uL verkhney vodnoy fazy v novye tubes obemom 1,5-2 mL.
4. Add 250 uL svyazyvayushchego bufera i peremeshat tubes s pomoshchyu mnogokratnogo perevorachivaniya.
5. Perenesti ves obem smesi v spin-kolonku.
6. Centrifuge tubes v techenie 1 minuty pri 10-13 tys. ob/min. Udalit filtrat iz sobiratelnykh probirok.
Promyvka RNA
1. Add v tubes 700 uL promyvochnogo bufera.
2. Centrifuge tubes v techenie 1 minuty pri 10-13 tys. ob/min. Udalit filtrat iz sobiratelnykh probirok.
3. Repeat punkty 1, 2.
4. Vernut kolonki v sobiratelnye tubes. Centrifuge tubes v techenie 1 minuty pri maks. ob/min.
Elyutsiya RNA
1. Perenesti spin-kolonki v novye tubes obemom 1,5-2 mL. Sobiratelnye tubes utilizirovat.
2. Add na tsentr membrany spin-kolonok 30-100 uL elyuiruyushchego bufera. Incubate at room temperature v techenie 3-5 minut. Rekomendatsiya: dlya predotvrashcheniya degradatsii RNA vnesti ingibitor RNAaz.
3. Centrifuge tubes pri +4℃ v techenie 1 minuty pri 13 tys. ob/min.
4. Utilizirovat spin-kolonki.
Poluchennye rastvory RNA gotovy k primeneniyu, vse dalneyshie manipulyatsii sleduet provodit pri +4℃. Po absorbance ratio A260/A280 purity of the resulting solution RNA sootvetstvuyut ≥1,9.
Pered khraneniem rekomenduetsya razdelit rastvory RNA na alikvoty, kotorye must be stored pri temperature -20℃ i nizhe ne bolee 1 goda. Dlya predotvrashcheniya degradatsii RNA rekomenduetsya dobavlyat ingibitor RNAaz i/ili khranit samples pri temperature -72°C.

Buffers nabora "RNA Pro Q" can be stored pri temperature ot +4 do +25℃ for 12 months from the manufacturer's release date. If a precipitate is present pri minimalnykh temperaturakh khraneniya, warm the solutions to room temperature.

Microcentrifuge tubes obemom 1,5-2 mL; shtativ dlya mikrotsentrifuzhnykh probirok obemom 1,5-2 mL; avtomaticheskie dozatory peremennogo obema na 20-200 uL, 100-1000 uL i sootvetstvuyushchie disposable tips; cpin-kolonki s sobiratelnymi probirkami (idut v komplekte); vortex mixer; skorostnaya tsentrifuga dlya probirok obemom 1,5-2 mL do 13 tys. ob/min s vozmozhnostyu podderzhaniya temperatury +4℃.