Reagent kit for bisulfite DNA modification "ConverGen" Raissol
Raissol
Reagent kit for bisulfite DNA modification "ConverGen" Raissol is a Raissol reagent product for ngs library preparation in molecular biology, genetics, diagnostic and research laboratories.
Application area
Used for sample preparation, nucleic acid analysis, PCR/qPCR workflows and sequencing-related projects where applicable.
Sample types
The product is intended for work with biological material.
Advantages of Raissol reagents
- stable reagent performance and reproducible laboratory results
- compatibility with common molecular biology protocols
- convenient workflow for routine and research procedures
- efficient preparation of biological material for downstream analysis
Key specifications
| Category | PCR reagents and supplies;Metilirovanie;Agrogenetics>>>Agrogenetics - library preparation;Raissol |
| Manufacturer | Raissol |
| Primary use | NGS library preparation |
| Sample type | biological material |
V nabor vklyucheny vse neobkhodimye reagenty:
1. Modifitsiruyushchiy reagent - sulfirovanie nemetilirovannykh ostatkov tsitozina;
2. Modifitsiruyushchiy rastvor 1 - prigotovlenie modifitsiruyushego reagenta;
3. Modifitsiruyushchiy rastvor 2 - prigotovlenie modifitsiruyushego reagenta;
4. Buffer dlya razvedeniya - razvedenie reagentov i obrazov DNA;
5. Binding buffer - dlya povysheniya sorbtsii NA na filtre kolonki;
6. Wash buffer - dlya promyvki nucleic acids;
7. Desulfiruyushchiy bufer - dlya udaleniya sulfogruppy;
8. Elution buffer - dlya elyutsii i khraneniya NA, pH=8,9;
9. Spin-kolonki - dlya sorbtsii NA.
1. Modifitsiruyushchiy reagent - sulfirovanie nemetilirovannykh ostatkov tsitozina;
2. Modifitsiruyushchiy rastvor 1 - prigotovlenie modifitsiruyushego reagenta;
3. Modifitsiruyushchiy rastvor 2 - prigotovlenie modifitsiruyushego reagenta;
4. Buffer dlya razvedeniya - razvedenie reagentov i obrazov DNA;
5. Binding buffer - dlya povysheniya sorbtsii NA na filtre kolonki;
6. Wash buffer - dlya promyvki nucleic acids;
7. Desulfiruyushchiy bufer - dlya udaleniya sulfogruppy;
8. Elution buffer - dlya elyutsii i khraneniya NA, pH=8,9;
9. Spin-kolonki - dlya sorbtsii NA.
Kratkaya instruktsiya "ConverGen"
Polnaya instruktsiya "ConverGen"
Prigotovlenie modifitsiruyushchego reagenta1. V tube s modifitsiruyushchim reagentom add 900 uL bufera dlya razvedeniya i 60 uL modifitsiruyushchego rastvora 1. Intensivno peremeshat tube on a vortex mixer, sbrosit kapli s pomoshchyu brief centrifugation.
2. Incubate tube v termostate pri 65°C v techenie 30 minut, periodicheski peremeshivaya on a vortex mixer.
3. Cbrosit kapli s pomoshchyu brief centrifugation.
4. Incubate tube at room temperature v techenie 3-5 minut until completely cooled.
5. Add v tube 200 uL modifitsiruyushchego rastvora 2. Intensivno peremeshat tube on a vortex mixer, sbrosit kapli s pomoshchyu brief centrifugation.
Primechanie: vozmozhno vypadenie osadka, sluchaynyy zakhvat kotorogo ne vliyaet na effektivnost bisulfitnoy modifikatsii.
Bisulfitnaya modifikatsiya DNA
1. V tubes obemom 200 uL vnesti 20 uL obraztsa DNA.
Primechanie: dlya bisulfitnoy modifikatsii mozhno ispolzovat ot 50 pg do 2 mkg DNA na reaktsiyu.
Optimalnaya kontsentratsiya obraztsa DNA sostavlyaet 20-50 ng/uL. Chistota obraztsa DNA po absorbance ratio A260/A280 dolzhna byt ≥1,7.
2. Add v tubes 120 uL modifitsiruyushchego reagenta. Mix tubes on a vortex mixer, sbrosit kapli s pomoshchyu brief centrifugation.
3. Pomestit tubes v amplifikator i zapustit sleduyushchuyu programmu:
Primechanie: modifitsirovannuyu DNA mozhno khranit pri +4°C v techenie 12 ch.
Desulfirovanie i ochistka DNA
Sorbtsiya NA
1. V spin-kolonki vnesti 400 uL svyazyvayushchego bufera i add ves obem obraztsa posle bisulfitnoy konversii. Mix smes s pomoshchyu perevorachivaniya spin-kolonok.
2. Centrifuge kolonki v techenie 30 sekund pri 10-12 tys. ob/min. Udalit filtrat iz sobiratelnykh probirok.
Desulfirovanie i promyvka NA
1. Add v kolonki 200 uL promyvochnogo bufera.
2. Centrifuge kolonki v techenie 30 sekund pri 10-12 tys. ob/min. Udalit filtrat iz sobiratelnykh probirok.
3. Add v kolonki 200 uL desulfiruyushchego bufera.
4. Incubate spin-kolonki at room temperature 20 minut.
5. Centrifuge kolonki v techenie 30 sekund pri 10-12 tys. ob/min. Udalit filtrat iz sobiratelnykh probirok.
6. Dvazhdy povtorit punkty 1-2.
7. Centrifuge pustye spin-kolonki v techenie 1 minuty pri 10-12 tys. ob/min.
8. Perenesti spin-kolonki v novye tubes obemom 1,5-2 mL. Sobiratelnye tubes utilizirovat.
Nucleic acid elution
1. Add na tsentr membrany spin-kolonok 22 uL elyuiruyushchego bufera. Incubate v techenie 5 minut at room temperature.
2. Centrifuge kolonki v techenie 1 minuty pri 10-12 tys. ob/min.
3. Utilizirovat spin-kolonki.
Optsionalno: dlya povysheniya vykhoda NA neobkhodimo povtorno nanesti filtrat na membranu kolonki.
Poluchennye rastvory modifitsirovannykh nucleic acids can be stored do 2 dney pri temperature ot +2°C do +4°C i do goda pri temperature -20°C.
Vse reagenty nabora "ConverGen" can be stored pri temperature ot +4°C do +25°C for 12 months from the manufacturer's release date. If a precipitate is present pri minimalnykh temperaturakh khraneniya, warm the solutions to room temperature.
Posle prigotovleniya modifitsiruyushchiy reagent sleduet khranit ne bolee mesyatsa pri temperature ot -20°C do -15°C.
Posle prigotovleniya modifitsiruyushchiy reagent sleduet khranit ne bolee mesyatsa pri temperature ot -20°C do -15°C.
Probirki obemom 0,2 mL; mikrotsentrifuzhnye tubes obemom 1,5-2 mL; shtativ dlya probirok obemom 0,2 mL; shtativ dlya mikrotsentrifuzhnykh probirok
obemom 1,5-2 mL; avtomaticheskie dozatory peremennogo obema na 2-20 uL, 20 200 uL, 100-1000 uL i sootvetstvuyushchie disposable tips; cpin-kolonki s sobiratelnymi probirkami (idut v komplekte); vortex mixer; tverdotelnyy termostat s vozmozhnostyu podderzhaniya temperaturnogo rezhima v diapazone 25-80oS dlya probirok obemom 1,5-2 mL; amplifikator; skorostnaya mikrotsentrifuga dlya probirok obemom 1,5-2 mL do 13 tys. ob/min.
obemom 1,5-2 mL; avtomaticheskie dozatory peremennogo obema na 2-20 uL, 20 200 uL, 100-1000 uL i sootvetstvuyushchie disposable tips; cpin-kolonki s sobiratelnymi probirkami (idut v komplekte); vortex mixer; tverdotelnyy termostat s vozmozhnostyu podderzhaniya temperaturnogo rezhima v diapazone 25-80oS dlya probirok obemom 1,5-2 mL; amplifikator; skorostnaya mikrotsentrifuga dlya probirok obemom 1,5-2 mL do 13 tys. ob/min.