Cells were collected in duplicates at 48 h and 9 days after doxycycline induction. We combined new iA RNA-seq with those published (Aydin et al., 2019 (link)) to make an n of 5. TRIzol (Invitrogen, 15596026) reagent was used to isolate RNA. Isolated RNA was purified with RNeasy mini kit (Qiagen, 74106). RNA integrity was measured using Agilent High Sensitivity RNA Screentape (Agilent Tech, 5067–5080). 500 ng RNA was spiked (1:100) with ERCC Exfold Spike-in mixes (Thermo Fisher Scientific, 4456739) for accurate comparison across samples. RNA-seq libraries were prepared with Illumina TruSeq LS kit v2 (RS-122–2001; RS-122–2002). KAPA library amplification kit was used for the final quantification of the library before pooling (Roche Lightcycler 480). The libraries were sequenced on an Illumina Next-Seq 500 using V2 and V2.5 chemistry for 50 cycles (single-end) at NYU Genomics Core facility.
Truseq ls kit v2
Manufactured by Illumina
The TruSeq LS kit v2 is a library preparation kit designed for the Illumina sequencing platform. The kit provides reagents and protocols for constructing sequencing libraries from DNA samples.
Automatically generated - may contain errors
Lab products found in correlation
Agilent high sensitivity rna screentape, by Agilent Technologies (3 mentions)
Ercc exfold spike in mixes, by Thermo Fisher Scientific (3 mentions)
Nextseq 500, by Illumina (3 mentions)
Rneasy mini kit, by Qiagen (3 mentions)
Kapa library amplification kit, by Roche (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
3 protocols using truseq ls kit v2
1
RNA-seq Analysis of Induced Cell Lines
2
RNA-seq analysis of Doxycycline induction
3
RNA-seq analysis of Doxycycline induction
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Cells were collected 0, 12 and 48 hours after Doxycycline induction and RNA was isolated by resuspending in Trizol reagent (Invitrogen, 15596026) followed by purification using Qiagen RNeasy mini kit (Qiagen, 74106). RNA integrity was measured with Agilent High Sensitivity RNA Screentape (Agilent Tech, 5067–5080). 500 ng of RNA was spiked-in (1:100) with ERCC Exfold Spike-in mixes (Thermo Fisher, 4456739) for accurate comparison across samples. Illumina TruSeq LS kit v2 (RS-122–2001; RS-122–2002) was used to prepare RNA-seq libraries. The final quantification of the library before pooling was done with KAPA library amplification kit (Roche Lightcycler 480). The libraries were sequenced on Illumina NextSeq 500 using V2 and V2.5 chemistry for 50 cycles (single-end) at the Genomics Core Facility at NYU.
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