English
Liza celic je pomembna eksperimentalna metoda za spremembo celične prepustnosti in aktivnosti. Pomeni, da potem, ko je celica okužena z virusom, virus uporabi hranila v gostiteljski celici za sintetiziranje lastnih potomcev, nato pa virusni potomci uničijo gostiteljsko celico in se sprostijo iz celic. V poskusih celične lize je bučke za celično kulturo običajen potrošni material. Koraki za celično lizo so naslednji:
1. Izlijte gojišče in stekleničko za celično kulturo privijte z glavo navzdol na vpojni papir, tako da bo vpojni papir absorbiral gojišče (ali postavite steklenico za nekaj časa pokonci, da preostanek gojišča lahko priteče na dno steklenice in ga nato s pipeto posesajte ).
2. V vsako steklenico celic dodajte 3 ml 4 ℃ predhodno ohlajenega PBS (0,01 M pH7,2～7,3). Celice položite ravno in jih nežno stresajte 1 minuto, da celice sperete, nato pa raztopino za pranje zavrzite. Zgornjo operacijo ponovite dvakrat, pri čemer trikrat sperete celice, da izperete kulturno tekočino. Zavrzite PBS in postavite bučko za kulturo na led.
3. Dodajte 10 ul PMSF (100 mM) v 1 ml pufra za lizo, dobro pretresite in postavite na led. (PMSF je treba pretresati, dokler ni kristalov, preden zmešate z raztopino za lizo.)
4. V vsako bučko celic dodajte 400 ul pufra za lizo, ki vsebuje PMSF, in lizirajte na ledu 30 minut. Za popolno liziranje celic je treba bučko pogosto stresati.
5. Po lizi s čistim strgalom postrgajte celice na eni strani bučke za kulturo (ukrep naj bo hiter), nato pa s pištolo premaknite celične ostanke in lizatirajte v 1,5-ml epruveto za centrifugiranje. (Celotna operacija naj poteka na ledu, kolikor je mogoče.)
6. Centrifugirajte pri 12000 vrt/min pri 4°C 5 minut. (Vnaprej vklopite centrifugo, da se predhodno ohladi)
7. Centrifugirani supernatant prenesite v centrifugalno epruveto za 0,5 min in ga shranite pri -20°C
bučke za celično kulturo se ne uporabljajo samo za poskuse celične lize, temveč tudi za srednje velike celične in tkivne kulture v laboratoriju, kot so celice Vero , celice HEK 293, celice CAR-T, celice CHO, celice žuželk itd.
The FAI climbed 5.9 percent year-on-year in the first 11 months of 2018, quickening from the 5.7-percent growth in Jan-Oct, the National Bureau of Statistics (NBS) said Friday in an online statement.
The key indicator of investment, dubbed a major growth driver, hit the bottom in August and has since started to rebound steadily.
In the face of emerging economic challenges home and abroad, China has stepped up efforts to stabilize investment, in particular rolling out measures to motivate private investors and channel funds into infrastructure.
Friday's data showed private investment, accounting for more than 60 percent of the total FAI, expanded by a brisk 8.7 percent.
NBS spokesperson Mao Shengyong said funds into weak economic links registered rapid increases as investment in environmental protection and agriculture jumped 42 percent and 12.5 percent respectively, much faster than the average.
In breakdown, investment in high-tech and equipment manufacturing remained vigorous with 16.1-percent and 11.6-percent increases respectively in the first 11 months. Infrastructure investment gained 3.7 percent, staying flat. Investment in property development rose 9.7 percent, also unchanged.