RNA isolation and protocol | 合法醫療器材資訊網

PreparetheRNAinthefollowingway:Add2µgofRNA(calculatedby2µg/theconcentrationinµg/µl)toasmallEppendorftube.Bringthetotalvolume ...

1.1 Using at least 106 cells, aspirate the media and wash once with ice-cold PBS (1–2 ml).

1.2 Aspirate the PBS (remove as much as possible) and add 1 ml TRIzol.

1.3 Scrape the plate briefly, then remove the TRIzol with a pipette and deposit the TRIzol/cell lysate into a 1.5 ml Eppendorf tube.

1.4 Leave at room temperature for 5 min.

1.5 Add 250 µl chloroform and shake the tube vigorously for about 15 sec.

1.6 Leave at room temperature for 5 min.

1.7 Centrifuge at 10,000 rpm for 5 min.

1.8 At this point, there will be three layers in each tube:

Top layer: clear, aqueous

Middle layer/interphase: white precipitated DNA

Bottom layer: pink organic phase

Carefully remove the aqueous phase using...