DNA extraction kit from blood plasma Cf Extra Raissol

The kit includes all reagents required for DNA extraction:
1. Lysis buffer - na osnove chaotropic agents i auxiliary components dlya optimal lysis;
2. Binding buffer - dlya sorbtsii NA na magnitnykh chastitsakh;
3. Resuspendiruyushchiy bufer - dlya razrusheniya konglomerata chastits;
4. Wash buffer 1 - dlya promyvki nucleic acids ot cell metabolites and salts;
5. Wash buffer 2 - dlya promyvki nucleic acids ot cell metabolites and salts;
6. Elution buffer - dlya elyutsii i khraneniya NA, pH=8,9;
7. Proteinase K - dlya bolee bystrogo i polnogo lizisa;
8. Magnitnye chastitsy - dlya sorbtsii NA.

DNA extraction procedure

Lysis
1. V tubes obemom 15 mL place 4,5 mL blood plasma.
2. Add 900 uL liziruyushchego bufera i 20 uL Proteinazy K.
3. Mix tubes s pomoshchyu perevorachivaniya.
4. Incubate tubes na vodyanoy bane pri 56-60℃ v techenie 60-90 minut, peremeshivaya kazhdye 10-20 minut.
5. Perenesti tubes na led i inkubirovat v techenie 2-3 minut. V sluchae otsutstviya okhladitelnykh elementov inkubirovat at room temperature v techenie 10-15 minut until completely cooled probirok.
Sorbtsiya NA
1. Add k smesi 9,6 mL svyazyvayushchego bufera i 130 uL magnitnykh chastits, khorosho peremeshat.
2. Incubate tubes at room temperature v techenie 30 minut, peremeshivaya kazhdye 5-10 minut, libo ustanovit na vrashchayushchiysya rotor s minimalnoy skorostyu.
Resuspendirovanie
1. Centrifuge tubes v techenie 5 minut pri maks. ob/min, posle chego udalit supernatant.
2. Add k kazhdomu osadku 850 uL resuspendiruyushchego bufera i razrushit konglomerat chastits s pomoshchyu pipetirovaniya.
3. Perenesti suspenziyu v novye tubes obemom 1,5-2 mL.
4. Incubate tubes na magnitnom shtative v techenie 5 minut.
5. Akkuratno, ne zadevaya pellet chastits, udalit supernatant.
Nucleic acid washing
1. Add v tubes 900 uL promyvochnogo bufera 1.
2. Intensivno peremeshat tubes on a vortex mixer, sbrosit kapli s pomoshchyu brief centrifugation.
3. Incubate tubes na magnitnom shtative v techenie 3 minut, posle chego akkuratno, ne zadevaya pellet chastits, udalit supernatant.
4. Repeat punkty 1-3.
5. Add v tubes 900 uL promyvochnogo bufera 2.
6. Repeat punkty 2,3.
7. Vysushit samples s otkrytymi kryshkami v termostate pri 56-60℃ v techenie 5 -10 minut libo at room temperature v techenie 20 minut until complete ispareniya promyvochnogo rastvora.
Nucleic acid elution
1. Add v tubes 50-100 uL elyuiruyushchego bufera.
2. Intensivno peremeshat tubes on a vortex mixer, sbrosit kapli s pomoshchyu brief centrifugation.
3. Incubate v termostate pri 56-60℃ v techenie 10 minut, periodicheski peremeshivaya. Cbrosit kapli s pomoshchyu brief centrifugation.
4. Incubate na magnitnom shtative v techenie 3 minut.
5. Perenesti elyuat v novye tubes obemom 1,5-2 mL.
Optsionalno: rekomenduetsya provesti dopolnitelnoe tsentrifugirovanie elyuata na maksimalnoy skorosti dlya udaleniya ostatochnogo kolichestva magnitnykh chastits.
Poluchennye rastvory nucleic acids can be stored do 7 dney pri temperature ot +2℃ do +4℃ i do dvukh let pri temperature -20℃. Pokazateli poluchennykh rastvorov DNA sootvetstvuyut diapazonu kontsentratsiy 0,3-1 ng/uL pri izmerenii s pomoshchyu nabora Spectra Q HS (RaissolTM, Rossiya) na pribore Qubit (Thermo Fisher Scientific, SShA). Spektr razmera ustanavlivaetsya s pomoshchyu elektroforeza na pribore Caliper (Perkinelmer, SShA). Standartnyy diapazon 160-200 bp

Download the short instruction "CFExtra" (pdf)

Download the full instruction "CFExtra" (pdf)

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Buffers nabora "CF Extra" 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.
Magnitnye chastitsy can be stored pri temperature ot +4℃ do +25℃ for 12 months from the manufacturer's release date. Pered applicationm suspenziyu chastits neobkhodimo tshchatelno vzbaltyvat do odnorodnogo sostoyaniya.
Enzyme Proteinase K khranitsya pri -20℃, shelf life is 12 mesyatsev. A short-term storage temperature increase is allowed (transportation) ot +4℃ do +25℃ for no more than 5 days.