DNA extraction kit from whole blood "Blood T" Raissol

The kit includes all reagents required for total DNA extraction:
1. RBC bufer - prednaznachen dlya udaleniya eritrotsitov krovi;
2. Lysis buffer - na osnove sodium lauryl sulfate i auxiliary components dlya optimal lysis;
3. Precipitation buffer 1 - dlya protein precipitation;
4. Precipitation buffer 2 - dlya precipitation NA;
5. Wash buffer 1 - dlya promyvki nucleic acids ot cell metabolites and salts;
6. Wash buffer 2 - dlya promyvki nucleic acids ot cell metabolites and salts;
7. Elution buffer - dlya elyutsii i khraneniya NA, pH=8,9;
8. Proteinase K - dlya bolee bystrogo i polnogo lizisa.

Total DNA extraction procedure

Lysis
1. V tubes obemom 1,5-2 mL place 300 uL krovi i add 900 uL RBC bufera.
2. Vstryakhnut tubes on a vortex mixer.
3. Incubate v techenie 5-10 minut at room temperature.
4. Centrifuge v techenie 1 minuty pri maks. ob/min (ne menee 13 tys. ob/min dlya nastolnykh tsentrifug), dalee udalit supernatant.
5. K poluchennomu osadku kletok add 300 uL liziruyushchego bufera i 20 uL Proteinazy K, shake vigorously on a vortex mixer until complete razrusheniya konglomerata kletok.
6. Incubate v termostate pri 55-60℃ v techenie 15-25 minut, periodicheski vstryakhivaya.
7. Perenesti tubes na led ili v kholodnyy shtativ i inkubirovat v techenie 1 minuty. V sluchae otsutstviya okhladitelnykh elementov inkubirovat at room temperature v techenie 5-10 minut until completely cooled smesi.
Nucleic acid sorption and precipitation
1. Add k smesi 50 uL osazhdayushchego bufera 1.
2. Intensivno peremeshat tubes on a vortex mixer, sbrosit kapli s pomoshchyu brief centrifugation.
3. Incubate v techenie 3-5 minut vo ldu ili v cooling rack. V sluchae otsutstviya okhladitelnykh elementov inkubirovat at room temperature v techenie 5-10 minut.
4. Centrifuge tubes v techenie 5 minut pri 13 tys. ob/min.
5. Akkuratno, ne zadevaya pellet, perenesti 300-330 uL supernatanta v novuyu tube obemom 1,5-2 mL.
6. Add 330 uL osazhdayushchego bufera 2 i peremeshat s pomoshchyu pyatikratnogo perevorachivaniya tubes.
7. Dlya uvelicheniya vykhoda DNA rekomenduetsya inkubirovat tubes v freezer pri -20℃ v techenie 25 minut.
8. Centrifuge tubes v techenie 5 minut pri 13 tys. ob/min, posle chego udalit supernatant.
Nucleic acid washing
1. Add v tubes 700 uL promyvochnogo bufera 1.
2. Intensivno peremeshat tubes on a vortex mixer.
3. Centrifuge tubes v techenie 5 minut pri 13 tys. ob/min, posle chego udalit supernatant.
4. Add v tubes 700 uL promyvochnogo bufera 2.
5. Repeat punkty 2,3.
6. Vysushit samples s otkrytymi kryshkami v termostate pri 42℃ ili v tsentrifuge-kontsentratore pri 30℃ until complete ispareniya promyvochnogo rastvora.
Nucleic acid elution
1. Add v tubes 50-100 uL elyuiruyushchego bufera.
2. Incubate v termostate pri 60℃ v techenie 5 minut, periodicheski peremeshivaya.
Poluchennye rastvory nucleic acids can be stored do 7 dney pri temperature ot +2℃ do +4℃ i do dvukh let pri temperature -20℃. Po absorbance ratio na A260/A280 purity of the resulting solution DNA sootvetstvuyut ≥1,7.

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Buffers nabora "GM Blood T" 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.
Enzyme Proteinase K must be stored pri -20℃, shelf life is 12 mesyatsev. A short-term storage temperature increase is allowed (transportation) fermenta Proteinase K ot +4℃ do +25℃ for no more than 5 days.