Txhawm rau kom tshem tawm cov xim ntawm cov chloroplasts raug tshem tawm hauv qhov tsis sib cais. Txhawm rau kom tshem tawm cov pigments los ntawm chloroplasts, peb yuav tsum tau tshem ntawm qhov tsis ua ntej. Dab tsi yog txoj hauv kev zoo tshaj los cais cov chloroplasts ntawm qhov tsis? siv lub centrifuge los cais qhov
Yuav ua li cas cais cov chloroplast?
Nyob rau hauv txoj kev cais chloroplast, cell phab ntsa tawg siv tshuab los yog homogenizer. Tom qab ntawd cov nplooj tsis tawg thiab cov khib nyiab ntawm tes raug tshem tawm los ntawm kev pom. Cov chloroplasts yog sau los ntawm centrifugation siv ib tug percoll gradient.
Koj ua li cas thiaj ua kom muaj chloroplast cais tsis tawm?
Npaj qhov xav tau ntawm 1X Chloroplast Isolation buffer los ntawm diluting 2X buffer Tshuag 1: 1 nrog dH2O dej Ntxiv 10 µL ntawm 10% BSA tov ib mL ntawm 1X Chloroplast Isolation Buffer. Ua ntej siv, ntxiv 1 µL ntawm DTT rau 1 mL ntawm Chloroplast Isolation Buffer. Sau npe qhov tsis pub dhau los ua 'Complete Buffer'.
Dab tsi yog nyob rau hauv ib qho kev sib cais ntawm chloroplast?
DNA cais tsis tawm muaj 100 mM NaCl, 100 mM Tris-HCl (pH 8.0), 50 mM EDTA, thiab 1 mM DTT Cov chloroplast lyse tau txais los ntawm incubating lub chloroplast pellet nrog 8 ml ntawm DNA cais tsis, 1.5 ml 20% SDS, 20 µl 2-Mercaptoethanol thiab 30 µl Proteinase K (10 mg / ml) rau hauv lub raj centrifuge ntawm 55 ° C rau 4 h.
Dab tsi yog pH ntawm qhov tsis siv los ua homogenize nplooj nplooj hauv chloroplast cais raws tu qauv?
Spinach yog ntim nrog cov chloroplasts zoo thiab lawv cais tau yooj yim. Koj yuav tsum tau muab cov nplooj tso rau hauv ib lub tshuab txias txias nrog 0.1 M phosphate tsis, pH 7.5 thiab 0.5 M sucrose kom tsis txhob tawg.
