Project description:The transcriptome profiling five tissues of juvenile Eriocheir sinensis, including gill, muscle, thoracic ganglion, eyestalk and hepatopancreas, were sequenced to get the basic dataset for constructed a genome-scale metabolic network model for E. sinensis. The model was used to predict the optimal nutrient requirements of E. sinensis in feed and suggestions for feed improvement were put forward based on the simulation results.
Project description:Total RNA extracted from the head and neck cell line CAL165 were purified using miRNeasy minikit (Qiagen). RNA libraries were then generated with the NEB next small library prep set for SOLID (New England Biolabs) and sequenced on the Applied Biosystems SOLiD 5500 wildfire system following the manufacturer's instructions.
Project description:Insect derived cell-lines, from Spodoptera frugiperda (Sf21) and from Trichoplusia ni (High Five), are ones of the most widely used systems for recombinant protein expression in Baculoviral Expression Vector System (BEVS). Genomic sequences and annotations are still incomplete for Sf21 or absent for High Five. In this study we present an approach using different sequencing data types with short-read sequencing, long synthetic and Oxford Nanopore reads to build genomes at an unprecedented resolution. The Sf21 and High Five assemblies contain 4,020 scaffolds of size, 463 Mb with N50 of 364 Kb and 2,954 scaffolds of size, 332 Mb with N50 of 326 Kb, respectively. Furthermore, we build a new gene prediction workflow, which integrates transcriptome proteome information using pre-existing tools. Using this approach, we could predict 21,506 Sf21 genes and 14,159 High Five genes, which were then functionally annotated. Finally, we also generate and integrate proteomic datasets to validate predicted genes. This integrative approach could be theoretically applied to any uncharacterized genome and result in valuable new resources. With this information available, Sf21 and High Five cells will become even better tools for protein expression and could be used in a wider range of applications, from promoter identifications to genome engineering and editing.
Project description:We sequenced cell-free RNA (cfRNA) for five cancer types (colorectal cancer, stomach cancer, liver cancer, lung cancer and esophageal cancer) and healthy individuals in 230 plasma samples collected from 6 clinical centers in China. Cancer related signaling pathway and microbial genus were identified. Cancer detection and specific classification were achieved through combining both host and microbial cfRNA reads.
Project description:Studying whether removal of base-excision repair from mitochondria will result into increase in mitochondrial DNA (mtDNA) mutation load. The endogenous genes of OGG1 and MUTYH DNA glycosylases were modified to lack the genomic region encoding for the predicted mitochondrial targeting sequence. The mouse lines used: A mouse line that lacks the region encoding for the mitochondrial targeting sequence (L2 to W23) of OGG1 (Ogg1 dMTS mice). A mouse line that lacks the region encoding the mitochondrial targeting sequence (K2 to P33) of MUTYH (Mutyh dMTS mice). To accumulate mutations to the mitochondrial DNA these mice were bred double homozygous Mutyh dMTS x Ogg1 dMTS mice as a maternal lineage for five consecutive generations and mitochondrial DNA from liver was extracted from the offspring and sequenced with Illumina. OGG1 and MUTYH are involved in repair of 8-oxo-dG from DNA. 8-oxo-dG can be a mutagenic lesion because some DNA repair polymerases are known to erroneously incorporate adenosine opposite to 8-oxo-dG during replication leading to GC>TA transversion mutations.
Project description:Bicyclus anynana butterflies were reared at 17°C and 27°C to produce the dry and wet season forms. RNA was extracted using TRIzol from the heads of 12 individual animals ~0-3 hours after eclosing; 3 dry season females, 3 wet season females, 3 dry season males, and 3 wet season males. A TruSeq RNA Sample Preparation Kit v2 was used to make 12 double stranded cDNA libraries from polyadenylated RNA. We size selected for DNA at ~280-340 bp. Libraries were sequenced using a HiSeq 2500, paired end 100-cycle sequence run.