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| | | Cell Culture Media, Kits, and Products Kultura stanica Media, Kits, i proizvodi | | |
Subculturing Cells - Cell Culture Subculturing Stanica - kultura stanica
Cell Culture Kultura stanica
Cell Subculture Protocol Stanica Subculture Protocol
SUBCULTURING CELLS SUBCULTURING stanice
Get DMEM media 4C; Trypsin-20C; BSA-20C; Get DMEM medija 4C; Trypsin-20C; BSA-20C; Put in 36mL of DMEM into conical tube plastic Put u 36mL, DMEM u konusna cijev plastični Add 4mL of BSA media to 36mL DMEM; Dodaj 4mL od nas mediji na 36mL DMEM; Get syringe and filter; Get šprica i filtar; Suck up media with syringe. Add filter. Sisati se medijima s šprica. Dodaj filtar. Push out media through filter on syringe. Push iz medija kroz filtar na brizgalicu. Repeat twice to get all of volume. Ponovite dva puta kako biste dobili sve prostore. Put into new plastic tube conical. Put u novi plastične cijevi konusna. Remove old media by dumping into beaker; DO NOT USE ASPIRATOR (get contamination!!) Uklonite stari mediji po damping u pehar, ne koristite usisivač (get kontaminacije!) Add 3mL typsin to culture. Trypsin with pull cells off the culture container. Dodaj 3mL typsin za kulturu. Trypsin s povuci stanica off kulture kontejner. Put container into incubator. Wait 2 min; Take out; look into microscope. Put kontejner u inkubator. Pričekajte 2 min; Take out; gledati u mikroskop. Try to get less clumps. More individual cells. Pokušajte dobiti manje razredi. Više pojedinih stanica. Remove most of trypsin by dumping into beaker. Ukloni većina trypsin po damping u pehar. Put back into incubator. Put nazad u inkubator. Check frequently. Provjerite često. Slap container on sides to check bottom for less clumps; look into microscope. Slap kontejner na dnu strane kako bi provjerili da za manje razredi; gledati u mikroskop. When see many individual cells floating along; Great! Kada se vidjeti mnoge pojedinačne stanice zajedno s pomičnim; Great! Add media to larger container; 1:5/ 1:3 dilution (3-5 containers larger). Dodaj medija za veće posuda; 1:5 / 1:3 dilution (3-5 posude većih). Add 10mL to (T75)large container, 5mL to (T25)small container. Dodaj 10mL (T75) veliki kontejner, 5mL (T25) mala posuda. Suck out Most (5mL of cells from T25). Sisati iz Mosta (5mL od stanica iz T25). Add to T75. Dodaj u T75. Leave some cells in small container. Add media to small container. Check in microscope for single cells; Incubate in 37C incubator. Ostavite neke stanice u malom kontejner. Dodaj medija u malom kontejner. Check in microscope za jednokrevetnu stanica; izleći piliće u 37C inkubator. See other Cell-protocols at our Cell Biology and Cell Culture Protocol Directory . Pogledajte druge Stanica-protokoli na naš stanična biologija i kulture stanica Protokol Directory.
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