| Accession | CRX151330 |
| Organism | Arabidopsis thaliana |
| Title | iSTARR-seq_plasmid_rep1 |
| BioProject | PRJCA003346 |
| BioSample | SAMC231922 |
| Platform | Illumina HiSeq X Ten |
| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
We extracted genomic DNA from two weeks Arabidopsis seedlings. About 30 µg of genomic DNA was diluted to 50ng/µl and fragmented by sonication (Scientz II-D). DNA fragments (500bp-800bp length) were size-selected, end repaired, 5’-phosphorylated and 3’ dA-tailed with VAHTS Mate Pair Library Prep Kit for Illumina® (VAHTS; cat. no. ND104). VAHTS Adapters for Illumina was ligated to about 6µg of DNA fragments using VAHTS Mate Pair Library Prep Kit for Illumina® (VAHTS; cat. no. ND104) following the manufacturer’s protocol. Adaptor ligated DNA was purified by using GeneJET PCR Purification Kit (Thermo Scientific; cat. no. K0702) then was amplified by TransStart FastPfu Fly DNA Polymerase (Transgen; cat. no. AP231) with Illumina sequencing primers (fw: ACACTCTTTCCCTACACGACG & rev: GACTGGAGTTCAGACGTGTGC). When the PolyA RNA was capture, keep the total RNA (without mRNA) supernatant, add 10ul RNase A TransGen® Biotech GE 101 20mg/ml at 37°C for 30-60min, and the plasmid was purified by GeneJET PCR Purification Kit and eluted in 50 ul EB. Purified plasmid as template for the second PCR (98°C for 2min; followed by 8-12 cycles of 98°C for 30s, 65°C for 30s, 72°C for 30s; and then 72 °C for 5min) with TransGen® Biotech FastPfu Fly DNA Polymerase and with VAHTSTM DNA Adapters for Illumina® (N302). PCR products were purified by GeneJET PCR Purification and eluted in 20-30 ul EB. All the libraries were sequenced on Illumine X Ten platform. |
OTHER |
GENOMIC |
PCR |
PAIRED
|
|
| Processing |
Planned read length (bp) for mate 1: 150
Planned read length (bp) for mate 2: 150
|
| Release date | 2020-10-14 |
|---|
| Run |
| Run accession |
Run data file information |
| File name | File size (MB) |
|---|
| CRR177596 |
CRR177596_f1.fq.gz
CRR177596_r2.fq.gz
|
1,384.26
1,537.35
|
| CRR177597 |
CRR177597_f1.fq.gz
CRR177597_r2.fq.gz
|
1,673.93
1,858.81
|
| CRR177598 |
CRR177598_f1.fq.gz
CRR177598_r2.fq.gz
|
1,644.32
1,827.75
|
| CRR177599 |
CRR177599_f1.fq.gz
CRR177599_r2.fq.gz
|
1,305.71
1,454.85
|
| CRR177600 |
CRR177600_f1.fq.gz
CRR177600_r2.fq.gz
|
1,665.74
1,843.88
|
| CRR177601 |
CRR177601_f1.fq.gz
CRR177601_r2.fq.gz
|
1,677.51
1,848.52
|
|
| Submitter | Jing Wan (1130014467@qq.com) |
|---|
| Organization | Huazhong Agricultural University |
|---|
| Date submitted | 2020-08-31 |
|---|
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