Frozen samples were ground into powders in liquid nitrogen with pestles and mortars. Total RNA was extracted using TRIzol (15596018; Invitrogen), and DNA was removed with the Turbo DNA-free Kit (AM1907; Invitrogen). The cDNA libraries were prepared from 1 µg RNA using the KAPA mRNA Hyper Prep Kit (KK8581; Roche) with the KAPA Dual-indexed Adapter Kit for Illumina platforms (KK8722; Roche). The libraries were sequenced with a NovaSeq 6000 (Illumina) at the Roy J. Carver Biotechnology Center at the University of Illinois at Urbana-Champaign.
Kapa dual indexed adapter kit for platforms
Manufactured by Illumina
The KAPA Dual-indexed Adapter Kit for Illumina platforms is a set of reagents designed to prepare DNA samples for sequencing on Illumina sequencing instruments. The kit provides adapter sequences that can be used to attach unique identifiers, or indexes, to individual DNA samples, enabling the pooling and subsequent identification of multiple samples in a single sequencing run.
Automatically generated - may contain errors
2 protocols using kapa dual indexed adapter kit for platforms
1
RNA Extraction and Sequencing Protocol
2
Transcriptome profiling of Arabidopsis seeds
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After 3 day of imbibition at 4 °C, WT seeds were sown on ½ MS medium and incubated in the growth chamber under constant light, 22 °C for 24 hours.
Seeds were transferred to ½ MS liquid medium supplemented with either 60 mM Glc, 5 µM ABA or both for 6 hours. Four biological replicates were collected for each treatment. Total RNA was extracted by RNeasy PowerPlant Kit (13500-50, Qiagen), DNA was removed via Turbo DNA-free kit (AM1907, Invitrogen). The cDNA libraries were prepared from 1000 ng RNA using KAPA mRNA Hyper Prep kit (KK8581, Roche) with KAPA Dual-indexed Adapter kit for Illumina platforms (KK8722, Roche). The libraries were sequenced by NovaSeq 6000 (Illumina) with paired-end reads at Roy J. Carver Biotechnology Center, UIUC.
Seeds were transferred to ½ MS liquid medium supplemented with either 60 mM Glc, 5 µM ABA or both for 6 hours. Four biological replicates were collected for each treatment. Total RNA was extracted by RNeasy PowerPlant Kit (13500-50, Qiagen), DNA was removed via Turbo DNA-free kit (AM1907, Invitrogen). The cDNA libraries were prepared from 1000 ng RNA using KAPA mRNA Hyper Prep kit (KK8581, Roche) with KAPA Dual-indexed Adapter kit for Illumina platforms (KK8722, Roche). The libraries were sequenced by NovaSeq 6000 (Illumina) with paired-end reads at Roy J. Carver Biotechnology Center, UIUC.
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