Chaf1a (s77588) and negative control Silencer Select siRNAs were purchased from LifeTechnologies. Dux siRNA pools were generated using Giardia Dicer. Briefly, primers were designed to amplify two ~400bp fragments of the endogenous Dux locus from genomic mouse DNA and add T7 handles (see Supplementary Table 14 ). Purified PCR products were then used as template for in vitro transcription using the MEGAscript® T7 Transcription Kit (ThermoFischer, AM1334). Template DNA was then degraded and the ssRNA allowed to anneal before dicing. Diced siRNAs were purified using the PureLink™ Micro-to-Midi Total RNA purification Kit (Invitrogen, 12183-018) with modifications. siRNA concentration was measured with the Qubit® RNA HS Assay Kit (ThermoFisher, Q32852). mESCs containing the MERVL:GFP reporter were transfected with 20pmol (10pmol of each) of total siRNA using RNAiMax (Life Technologies). All siRNA transfections were performed twice (on back to back days) to ensure knockdown.
Chaf1a s77588
Manufactured by Thermo Fisher Scientific
Chaf1a (s77588) is a lab equipment product from Thermo Fisher Scientific. It is a specialized tool used for scientific research and analysis purposes. The core function of this product is to perform specific tasks related to the intended application, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Silencer select sirna, by Thermo Fisher Scientific (2 mentions)
Qubit rna hs assay kit, by Thermo Fisher Scientific (2 mentions)
Purelink micro to midi total rna purification kit, by Thermo Fisher Scientific (2 mentions)
Megascript t7 transcription kit, by Thermo Fisher Scientific (2 mentions)
Rnaimax, by Thermo Fisher Scientific (2 mentions)
2 protocols using chaf1a s77588
1
Knockdown of Dux in mESCs
2
Knockdown of Dux in mESCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Chaf1a (s77588) and negative control Silencer Select siRNAs were purchased from LifeTechnologies. Dux siRNA pools were generated using Giardia Dicer. Briefly, primers were designed to amplify two ~400bp fragments of the endogenous Dux locus from genomic mouse DNA and add T7 handles (see Supplementary Table 14 ). Purified PCR products were then used as template for in vitro transcription using the MEGAscript® T7 Transcription Kit (ThermoFischer, AM1334). Template DNA was then degraded and the ssRNA allowed to anneal before dicing. Diced siRNAs were purified using the PureLink™ Micro-to-Midi Total RNA purification Kit (Invitrogen, 12183-018) with modifications. siRNA concentration was measured with the Qubit® RNA HS Assay Kit (ThermoFisher, Q32852). mESCs containing the MERVL:GFP reporter were transfected with 20pmol (10pmol of each) of total siRNA using RNAiMax (Life Technologies). All siRNA transfections were performed twice (on back to back days) to ensure knockdown.
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