U2OS-EJ2-GFP cells with MMEJ reporter24 (link) (1 × 105) were plated on 6 well plate and transfected 24 hr later with 2.5 µg pCMV-3xNLS-I-SceI or 0.5 µg control vector pCMV-3xNLS using Lipofectamine 2000 (Invitrogen). GFP+ frequencies were measured 3 days post transfection by FACS using GUAVA flow cytometer (Millipore) in triplicates and corrected for transfection efficiency and background events. Transfection efficiency was measured simultaneously by parallel transfection with 0.05 µg wt GFP expression vector (pCMV-3xNLS-GFP). For siRNA experiments, cells were transfected with 100 ρmol siRNA (Polθ SMARTpool (Dharmacon) or scrambled control) + 0.3 µg of pCMV-3xNLS-I-SceI (or control vector, GFP expression vector) per well.
Polθ smartpool
Manufactured by Horizon Discovery
The Polθ SMARTpool is a collection of pre-designed small interfering RNAs (siRNAs) targeting the DNA polymerase theta (Polθ) gene. The Polθ SMARTpool is designed to effectively silence the expression of the Polθ gene, which is involved in DNA repair processes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using polθ smartpool
1
MMEJ Repair Assay in U2OS Cells
2
MMEJ Repair Assay in U2OS Cells
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