siRNA introduction to Jurkat

Cell Information

Cell condition: 0.5`1.0~107cells/0.5mL medium
Medium: RPMI1640+10%FBS

Protocol

Reagent used: GenomONE
Trangient or Stable: Transient
Introduced molecule: Texas Red-labeled siRNA
Molecule conc.: 0.3g/L
Method: Method-2
Reagent volume: <Incorporation step>
HVJ-E: 40L
siRNA sol.: 10L
Reagent A: ---
Buffer: 20L
Reagent B: 3L
<Introduction step>
Reagent C: 5L
Buffer: 30L
Treatment/well:(*) 35L/well of 24-well plate
(*)
Add 35L into 0.5`1.0~107cells/0.5mL medium in micro test tube and centrifuge 1000g~30min at 4.
After centrifuging, remove supernatant and mix with 0.5mL of medium on 1 well of 24-well plate.
Incubation time: 48hr

Result

Efficiency: 90%@

.

Reference: Tomonori Ishii, Kei Ohnuma et al. :
Journal of Immunology, 170 (1), 3653-3661 (2003)

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