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Scriptseq 2

Manufactured by Illumina
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The ScriptSeq 2 is a library preparation kit designed for RNA sequencing. It enables the generation of cDNA libraries from RNA samples for downstream sequencing applications.

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Lab products found in correlation

A3500, by Promega (1 mentions) Truseq 2, by Illumina (1 mentions) Sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions) Single cell rna purification kit, by Norgen Biotek (1 mentions) Ribo zero magnetic kit, by Illumina (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Hiseq 2500, by Illumina (1 mentions)

Close spelling variants of this product

ScriptSeq version 2 RNA-seq library preparation kit Epicentre ScriptSeq v 2 RNA-seq library preparation kit ScriptSeq 2.0 kit

2 protocols using scriptseq 2

1

Single-Cell RNA Extraction and RT-qPCR Analysis

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was extracted using the Norgen single-cell RNA purification kit (Norgen, 51800) with on-column DNase digestion (Norgen, 25710). Four-hundred nanograms of total RNA was transcribed to cDNA using the reverse transcription system (Promega, A3500) and random primers. qPCRs were performed using the SYBR Green PCR master mix (Applied Biosystems, 4309155) with standard cycling conditions. Unless indicated otherwise, data were normalized to Gapdh expression levels.
For splice isoform RT-qPCRs, the specificity of PCR products was analyzed by sequencing of PCR products. PSI values were calculated as described previously (Han et al. 2013 (link); Camacho Londoño and Philipp 2016 (link)) by the following formula:
PSI=100×2−dCT(Exon)2−dCT(Exon)+2−dCT(Exonskip).
qPCR primer sequences are in Supplemental Table S11.
Sequencing library preparation was performed using either ScriptSeq 2 (Epicentre, SSV21106) or TruSeq 2 (Illumina, RS-122-2001) kits.
Welte T., Tuck A.C., Papasaikas P., Carl S.H., Flemr M., Knuckles P., Rankova A., Bühler M, & Großhans H. (2019). The RNA hairpin binder TRIM71 modulates alternative splicing by repressing MBNL1. Genes & Development, 33(17-18), 1221-1235.
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2

Transcriptional profiling of RONA colonies

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Two biological replicates of RONA colonies were dissected away from supporting cells, disassociated and plated. Twenty-four hours after plating, total RNA was harvested from culture using Trizol (Invitrogen) following the manufacturer’s protocol. Ribosomal RNAs were subtracted from total RNA using RiboZero magnetic kit (Epicentre). Libraries from rRNA-subtracted RNA were generated using ScriptSeq 2 (Epicentre) and sequenced on an Illumina HiSeq 2500. Reads were mapped to the human genome (release 19) using TopHat2. Reads were annotated using a custom R script using the UCSC known Genes table as a reference. Gene ontology shows output from a enrichment analysis on DAVID (david.abcc.ncifcrf.gov/), searching the following categories: GO Biological Process, GO Molecular Function, PANTHER Biological Process, PANTHER Molecular Function, KEGG and PANTHER pathway. Enrichment terms and associated genes and statistics are presenting with additional statistical analyses also provided. P-values presented in the text are corrected for multiple comparisons using the Benjamini method (column L).
Xu J.C., Fan J., Wang X., Eacker S.M., Kam T.I., Chen L., Yin X., Zhu J., Chi Z., Jiang H., Chen R., Dawson T.M, & Dawson V.L. (2016). Cultured networks of excitatory projection neurons and inhibitory interneurons for studying human cortical neurotoxicity. Science translational medicine, 8(333), 333ra48.
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