A method of measuring the concentration of an siRNA by single molecule fluorometry comprising; the first step wherein, in a test solution containing a nucleic acid extract prepared from a biological sample to which the siRNA has been administered, dissociating the double strands of the siRNA and ...
the cells were transfected 24 hours after plating, using 2 µl siport lipid (ambion) according to the manufacturer's protocol, at a final sirna concentration of 75 nm. immunofluorescence analysis was performed 96 hr post transfection using mouse anti-human ß-actin prim 9、ary antibody and...
Accell siRNA retains functionality following storage at a variety of conditions. Accell siRNA targeting human GAPD and Accell Non-targeting control siRNA (NTC) were stored at 1 mM concentration in RNAse-free water (10 μM data not shown) 由于Accell siRNA最终需要混合到培养基中,实验结果表明,加入...
that when you purchase three Silencer Pre-designed siRNAs to the same target, at least two of the siRNAs will reduce target mRNA levels in cultured cells by 70% or more when measured 48 hours after transfection at 100 nM or higher final siRNA concentration under the conditions described below...
Fig.1. Superior knockdown with RFectPM transfection reagent on difficult transfection primary cells. Different primary cells were transfected with scrambled or Lamin A/C siRNA (final concentration: 10nM) using RFectPM .In this and all the following figures, all the cells are harvested 48 hours after...
Use 10 nM siRNA as a starting point, although siRNA concentrationcan be much lower than 10 nM. Forward transfection and reverse transfection protocols can be usedfor most cell lines.Protocol: Forward TransfectionUse the following procedure to transfect siRNA into cells in a 24-wellformat...
knockdown levels despite differences in cell density, minor experimental inaccuracies, and other variations. Not also does this make it easy to optimize Lipofectamine RNAiMAX reagent for the lowest siRNA concentration but also reduces cellular stress and cytotoxicity in your experimental system ...
(a) Viability of Panc-1 cells after 24 h incubation of GNC–siRNA versus the identical siRNA concentration coupled with Lipofectamine 2000 transfection agent (Lipo2000-siRNA). Mean±s.d. (n ¼ 3). (b) Cells were pre-incubated with GNC–siRNA or GNC–nsRNA (100 nM siRNA equivalent) ...
转染实验优化: 为了提高转染效率,最好对转染条件进行优化,特别是首次使用。例如:24孔培养板,调整 siRNA与RFectPM试剂的用量。siRNA用量在0.6-30 pmol (final concentration 1- 50 nM)之间调整,RFectPM试剂用量在1.0 - 3.0μl之间调整。 原代细胞转染实验要点: ...
3siRNAconcentrationgradientgroups(50、100、200nmol/L)have been set印.24 hoursafter transfection,apoptosis wasexamined by FCM essay.RT-PCR wasusedtoexaminethe expression ofsurvivinmRNAand Immunocytochemistry was usedtoevaluatethe expression ofsurvivin ...