|
| Status |
Public on Sep 27, 2023 |
| Title |
Callus RNA-seq rep2 |
| Sample type |
SRA |
| |
|
| Source name |
leaf
|
| Organism |
Oryza sativa |
| Characteristics |
ecotype: japonica
developmental stage: Callus
|
| Treatment protocol |
Calli were treated with different combinations of transformation treatments.
|
| Growth protocol |
Plants derived derectly from seeds or regenerated from calli were cultivated until the tillering stage in an Academia Sinica greenhouse under natural light.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated from leaves of transgenic rice under different treatment groups using RNeasy Plant mini kit (Qiagen) or Trizol for RNA-seq libraries. Genomic DNA was extracted from leaves by using a DNeasy Plant Mini Kit (Qiagen).
RNA-seq libraries were prepared following standard Illumina protocols according to manufacturer’s instructions. The fragmented DNA was then used for library preparation using Illumina's library preparation kits. Premethylated adapter ligated fragments were bisulfite converted using the EZ DNA methylation kit (Zymo Research). The BS-seq libraries were then subjected to PCR amplification prior to sequencing on the Illumina HiSeq 2500 (Illumina) sequencers.
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina HiSeq 2500 |
| |
|
| Description |
Callus_2
|
| Data processing |
Clean RNA-seq reads were mapped to the genome using Tophat.
Fragments per kilobase of transcript per million mapped reads (FPKM) was obtained to represent the gene expression level. Count table was generated using Cuffnorm and DEGs were identified using Cuffdiff. Genes with adjusted p-values <=0.05 and 1.5-fold difference in expression were considered differentially expressed.
BS-seq reads were aligned against the rice genome (MSU7) using BS Seeker2
BS Seeker2 post-processes the alignments to remove non-unique and low-quality mappings.
The DNA methylation level was calculated as ((#C)/(#C+#T)), each of which covered at least four reads for an accurate estimation
Assembly: MSU7
Supplementary files format and content: Processed data (CGmap) is in text format. column 1=chromosome id; column 2=C/G (G stands for C on the opposite strand); column 3= chromosomal location; column 4=sequence context (CG/CHG/CHH); column 5=sequence context (CA/CT/CC/CG); column 6=methylation fraction #C/(#C+#T); column 7=number of C (#C); column 8 #C+#T.
|
| |
|
| Submission date |
May 11, 2022 |
| Last update date |
Sep 27, 2023 |
| Contact name |
Ying-Chung Lin |
| E-mail(s) |
ycjimmylin@ntu.edu.tw
|
| Organization name |
National Taiwan University
|
| Department |
Life Science
|
| Street address |
NO. 1, SEC. 4, ROOSEVELT RD., DA-AN DIST.
|
| City |
Taipei |
| ZIP/Postal code |
10617 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL19290 |
| Series (1) |
| GSE202715 |
Dissecting the Dynamic Changes of Methylome and Transcriptome During Rice Transformation |
|
| Relations |
| BioSample |
SAMN28189784 |
| SRA |
SRX15232105 |
