siRNA transfection with HiPerfect (MIN6 cells)


Posted by gregku
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Protocol

How to transfect MIN6 cells with siRNAs using HiPerfect.

Methods: 

1) Mix 5 pmoles of siRNA into 24.25 uL of Optimem (CCF or Invitrogen) in single well of 96 well plate. Add 0.75 uL HiPerfect (Qiagen). Incubate at RT 5 minutes (I have waited as long as 30 minutes).

2) Trypsinize MIN6 cells. Ideally, they should be in log phase (0.7-1.0 million per ml). Place 5000 cells in 175 uL of complete media and add to 96 well containing the siRNA and HiPerfect.

3) Analyze 3-5 days later.

Notes: For 384 well plates, I use 1.25 pmoles siRNA in 9.5 uL of Optimem, 0.5 uL of HiPerfect, 40 uL of cells. I do recommend testing different siRNA concentrations and HiPerfect amounts for your particular cell line.