DNA extraction kit from amniotic fluid "Amex" Raissol

The kit includes all reagents required for DNA extraction:

1. Lysis buffer - na osnove sodium lauryl sulfate i auxiliary components dlya optimal lysis;
2. Precipitation buffer 1 - dlya protein precipitation;
3. Precipitation buffer 2 - dlya precipitation NA;
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.

Download the short instruction "Amex" (pdf)

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Lysis
1. V tubes obemom 1,5-2 mL place 1,5-2 mL amniotic fluid.
2. Centrifuge tubes v techenie 3 minut pri 13 tys. ob/min, dalee, ne zadevaya pellet, udalit supernatant.
3. K poluchennomu osadku add 1,5-2 mL amniotic fluid.
4. Repeat punkt 2.
5. K poluchennomu osadku kletok add 300 uL liziruyushchego bufera i 15 uL Proteinazy K, shake vigorously on a vortex mixer until complete razrusheniya konglomerata kletok.
6. Incubate v termostate pri 56-60℃ v techenie 25 minut, periodicheski vstryakhivaya.
7. Cbrosit kapli s pomoshchyu brief centrifugation.
8. Pomestit 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 probirok.

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 on ice ili v cooling rack, periodicheski peremeshivaya samples. 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 mnogokratnogo perevorachivaniya tubes.
Optsionalno: dlya uvelicheniya vykhoda DNA rekomenduetsya inkubirovat tubes v freezer pri -20℃ ne menee 25 minut.
7. 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 56-60℃ v techenie 5 minut, periodicheski peremeshivaya on a vortex mixer.
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 corresponds to ≥1,7.

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Buffers nabora "Amex" 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.