Protocol tips
| Upstream tips |
Protocol tips |
Downstream tips |
|
siDKC1 #1. Custom-designed ON-TARGETplus SMARTpool siRNA pool targeting only
the 3’-untranslated region
a. GGAUAUGGGUGGUGAAAGA
b. CCUCAAAGCUUGUGUACAG
c. CUGAGAGAGUGGAACAUAG
d. GCUAGUGGUUGUCUUAACA
siDKC1 #2. Pre-designed ON-TARGETplus SMARTpool siRNA pool targeting only the
coding region
a. CAAGGUGACUGGUUGUUUA
b. GCAGGUAGUUGCCGAAGCA
c. UCUCAUAAACGGCUGGUUA
d. GGACAGGUUUCAUUAAUCU
Add diluted siRNA to diluted Lipofectamine Reagent
Incubate for 5 min at RT and add to cells.
Incubate cells for 1–3 days at 37°C |
Analyse cells after 48 and 72h |
| Protocol tips |
siDKC1 #1. Custom-designed ON-TARGETplus SMARTpool siRNA pool targeting only
the 3’-untranslated region
a. GGAUAUGGGUGGUGAAAGA
b. CCUCAAAGCUUGUGUACAG
c. CUGAGAGAGUGGAACAUAG
d. GCUAGUGGUUGUCUUAACA
siDKC1 #2. Pre-designed ON-TARGETplus SMARTpool siRNA pool targeting only the
coding region
a. CAAGGUGACUGGUUGUUUA
b. GCAGGUAGUUGCCGAAGCA
c. UCUCAUAAACGGCUGGUUA
d. GGACAGGUUUCAUUAAUCU
Add diluted siRNA to diluted Lipofectamine Reagent
Incubate for 5 min at RT and add to cells.
Incubate cells for 1–3 days at 37°C |
| Downstream tips |
| Analyse cells after 48 and 72h |
Publication protocol
Custom-designed (#1) and pre-designed (#2) ON-TARGETplus SMARTPool siRNA duplexes targeting DKC1 and negative control, ON-TARGETplus Non-Targeting siRNA pools #1 and #2 were obtained from Dharmacon (Lafayette, CO). The siRNA sequences are listed in Table S1. Transfections were performed using Lipofectamine 2000 (Invitrogen), as previously described [6].
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Manufacturer protocol
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