Nucleic acid extraction kit from milk "Meta Milk Maxi" 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 povysheniya sorbtsii NA na magnitnykh chastitsakh;
3. Resuspendiruyushchiy bufer - dlya razrusheniya konglomerata chastits;
4. Wash buffer 1 - dlya promyvki nucleic acids ot organicheskikh primesey i soley;
5. Wash buffer 2 - dlya promyvki nucleic acids ot organicheskikh primesey i soley;
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 3 mL issleduemykh prob milk.
2. Add k obraztsam 1 mL liziruyushchego bufera i 10 uL Proteinazy K.
3. Mix tubes s pomoshchyu perevorachivaniya.
4. Incubate tubes na vodyanoy bane pri temperature 56-60℃ v techenie 40 minut, periodicheski peremeshivaya.
Sorbtsiya NA
1. Add v tubes 8 mL svyazyvayushchego bufera i 50 uL magnitnykh chastits, peremeshat.
2. Incubate tubes at room temperature v techenie 20-30 minut, peremeshivaya kazhdye 5 minut, libo ustanovit na vrashchayushchiysya rotor s minimalnoy skorostyu.
3. Centrifuge tubes v techenie 5 minut pri maks. ob/min, posle chego udalit supernatant.
Resuspendirovanie
1. Add k kazhdomu osadku 500 uL resuspendiruyushchego bufera i razrushit konglomerat chastits s pomoshchyu pipetirovaniya.
2. Perenesti suspenziyu v novye tubes obemom 1,5-2 mL.
3. Incubate tubes na magnitnom shtative v techenie 5 minut.
4. Akkuratno, ne zadevaya pellet chastits, udalit supernatant.
Nucleic acid washing
1. Add v tubes 500 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 500 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 until complete ispareniya promyvochnogo rastvora.
Nucleic acid elution
1. Add v tubes 40-60 uL elyuiruyushchego bufera.
2. Intensivno peremeshat tubes on a vortex mixer, sbrosit kapli s pomoshchyu brief centrifugation.
3. Incubate v termostate pri temperature 56-60℃ v techenie 5 minut, periodicheski vstryakhivaya.
4. Sbrosit kapli s pomoshchyu brief centrifugation.
5. Incubate tubes na magnitnom shtative v techenie 3-5 minut.
6. Akkuratno, ne zadevaya chastitsy, perenesti elyuat v novye tubes.
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℃. Po absorbance ratio na A260/A280 purity of the resulting solution DNA corresponds to ≥1,7.

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Buffers nabora "Meta Milk Maxi" 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 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.