| Accession | CRX288849 |
| Organism | Triticum aestivum |
| Title | SPL13D |
| BioProject | PRJCA007017 |
| BioSample | SAMC476043 |
| Platform | Illumina NovaSeq 6000 |
| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
Full length TF was cloned into pIX-Halo vector, the HALO-SPL protein was generated using 500 ng of pIX-HALO-SPL plasmid and the TNT SP6 Coupled Reticulocyte Lysate System (Promega, L4600) according to the manufacturer’s protocol. The reaction was then incubated with 10 μL of Magne-HALO Tag beads (Promega, G7282) for 1 h at 25 °C in 1× PBS with 0.005% Nonidet P-40 (PBST). Bound protein was washed five times with PBST and subsequently treated with DNaseI. The DNA-free HALO-SPL protein was incubated with 500 μg of ultrasonicated genomic DNA library (200~800 bp) for 1 h at 25 °C. The beads were then washed eight times with PBST and incubated with 1μl Tn5 transposomes in 40 μl of tagmentation buffer (10 mM TAPS-NaOH ph 8.0, 5 mM MgCl2) at 55°C for 10 minutes. The DNA were recovered by NEB Monarch™ DNA Cleanup Kit (T1030L) and then amplified using Phusion DNA polymerase for 10-34 cycles. Amplified libraries were purified with AMPure beads to remove primers. |
OTHER |
GENOMIC |
PCR |
PAIRED
|
|
| Processing |
Planned read length (bp) for mate 1: 150
Planned read length (bp) for mate 2: 150
Insert size (bp): 400
|
| Release date | 2022-12-31 |
|---|
| Run |
| Run accession |
Run data file information |
| File name | File size (MB) |
|---|
| CRR336713 |
CRR336713_f1.fastq.gz
CRR336713_r2.fastq.gz
|
6,504.26
5,237.12
|
|
| Submitter | Zefu Lu (luzefu@caas.cn) |
|---|
| Organization | Institute of Crop Sciences, Chinese Academy of Agricultural Sciences |
|---|
| Date submitted | 2021-11-02 |
|---|
