Preparation Kit

產品描述：

1.      DNA起始量10ng-500ng，200ng-1ug可進行PCR free建庫；

2.      沒有GC偏好性，無接頭二聚體；

3.      具有384個dual index及384個單端index；

4.      除接頭外只有3管試劑（混勻mix），操作簡單；

試劑盒文獻：

1.      A male-expressed rice embryogenic trigger redirected for asexual propagation through seeds

1 January, 2019

Khanday I, Skinner D, Yang B, Mercier R, Sundaresan V

The molecular pathways that trigger the initiation of embryogenesis after fertilization in flowering plants, and prevent its occurrence without fertilization, are not well understood1. Here we show in rice (Oryza sativa) that BABY BOOM1 (BBM1), a member of the AP2 family2 of transcription factors that is expressed in sperm cells, has a key role in this process. Ectopic expression of BBM1 in the egg cell is sufficient for parthenogenesis, which indicates that a single wild-type gene can bypass the fertilization checkpoint in the female gamete. Zygotic expression of BBM1 is initially specific to the male allele but is subsequently biparental, and this is consistent with its observed auto-activation. Triple knockout of the genes BBM1, BBM2 and BBM3 causes embryo arrest and abortion, which are fully rescued by male-transmitted BBM1. These findings suggest that the requirement for fertilization in embryogenesis is mediated by male-genome transmission of pluripotency factors. When genome editing to substitute mitosis for meiosis (MiMe)3,4 is combined with the expression of BBM1 in the egg cell, clonal progeny can be obtained that retain genome-wide parental heterozygosity. The synthetic asexual-propagation trait is heritable through multiple generations of clones. Hybrid crops provide increased yields that cannot be maintained by their progeny owing to genetic segregation. This work establishes the feasibility of asexual reproduction in crops, and could enable the maintenance of hybrids clonally through seed propagation5,6.

Pages: 91-95   doi: 10.1038/s41586-018-0785-8

2.      Improved genomic resources for the black tiger prawn (Penaeus monodon)

4 February, 2020

Van Quyen D, Gan HM, Lee YP, Nguyen DD, Nguyen TH, Tran XT, Nguyen VS, Khang DD, Austin CM

World production of farmed crustaceans was 7.8 million tons in 2016. While only making up approximately 10% of world aquaculture production, crustaceans are generally high-value species and can earn significant export income for producing countries. Viet Nam is a major seafood producing country earning USD 7.3 billion in 2016 in export income with shrimp as a major commodity. However, there is a general lack of genomic resources available for shrimp species, which is challenging to obtain due to the need to deal with large repetitive genomes, which characterize many decapod crustaceans. The first tiger prawn (P. monodon) genome assembly was assembled in 2016 using the standard Illumina PCR-based pair-end reads and a computationally-efficient but relatively suboptimal assembler, SOAPdenovo v2. As a result, the current P. monodon draft genome is highly fragmented (> 2 million scaffolds with N50 length of &lt;1000 bp), exhibiting only moderate genome completeness (< 35% BUSCO complete single-copy genes). We sought to improve upon the recently published P. monodon genome assembly and completeness by generating Illumina PCR-free pair-end sequencing reads to eliminate genomic gaps associated with PCR-bias and performing de novo assembly using the updated MaSurCA de novo assembler. Furthermore, we scaffolded the assembly with low coverage Nanopore long reads and several recently published deep Illumina transcriptome paired-end sequencing data, producing a final genome assembly of 1.6 Gbp (1,211,364 scaffolds; N50 length of 1982 bp) with an Arthropod BUSCO completeness of 96.8%. Compared to the previously published P. monodon genome assembly from China (NCBI Accession Code: NIUS01), this represents an almost 20% increase in the overall BUSCO genome completeness that now consists of more than 90% of Arthropod BUSCO single-copy genes. The revised P. monodon genome assembly (NCBI Accession Code: VIGR01) will be a valuable resource to support ongoing functional genomics and molecular-based breeding studies in Vietnam.

Pages: 100751   doi: 10.1016/j.margen.2020.100751

3.      Sequence variation among SARS-CoV-2 isolates in Taiwan

1 January, 2020

Gong YN, Tsao KC, Hsiao MJ, Huang CG, Huang PN

Taiwan experienced two waves of imported cases of coronavirus disease 2019 (COVID-19), first from China in January to late February, followed by those from other countries starting in early March. Additionally, several cases could not be traced to any imported cases and were suspected as sporadic local transmission. Twelve full viral genomes were determined in this study by Illumina sequencing either from virus isolates or directly from specimens, among which 5 originated from clustered infections. Phylogenetic tree analysis revealed that these sequences were in different clades, indicating that no major strain has been circulating in Taiwan. A deletion in open reading frame 8 was found in one isolate. Only a 4-nucleotide difference was observed among the 5 genomes from clustered infections.

  doi: 10.1101/2020.03.29.014290v1.abstract

4.      Whole genome assembly of the snout otter clam, Lutraria rhynchaena, using Nanopore and Illumina data, benchmarked against bivalve genome assemblies

22 October, 2019

Thai BT, Lee YP, Gan HM, Austin CM, Croft LJ

Production of cultured bivalve molluscs was 17.1 million tons in 2016 accounting for 21.4% of global aquaculture production (FAO, 2018). The lack of genomic resources coupled with limited understanding of the molecular basis of gene expression and phenotypic variation have limited advances in aquaculture-based productivity of marine bivalves. Understanding the molecular basis of phenotypic variation and gene function is therefore important for selective breeding programs for traits such as increased growth and disease resistance. Similarly, whole genome assembles support GWAS studies to identify trait-specific loci and for genomic-based selective breeding. To this end, whole-genome sequencing has been conducted on several commercial bivalve species, including the edible oysters Crassostrea virginica (Gómez-Chiarri et al., 2015), Crassostrea gigas (Gerdol et al., 2015), pearl oysters (Takeuchi et al., 2012) and clams (Mun et al., 2017). However, in general, genomic data for bivalve molluscs, which includes a taxonomically diverse group of species, is sparse (Takeuchi et al., 2012; Zhang et al., 2012; Murgarella et al., 2016; Du et al., 2017; Li et al., 2017; Mun et al., 2017; Sun et al., 2017; Uliano-Silva et al., 2017; Wang et al., 2017; Li et al., 2018; Powell et al., 2018; Renaut et al., 2018; Bai et al., 2019). In this study we present the first genomic resources for a species of clam from the superfamily Mactroidea and for a Vietnamese shellfish species and generate a draft reference genome to form the basis of on-going selective breeding studies. This study also demonstrates the efficacy of using Oxford Nanopore Technology (ONT) reads to scaffold a bivalve genome assembly and shows the value of these relatively inexpensive long reads for spanning large repetitive regions and overcoming complex assembly issues caused by high heterozygosity, which typically confounds short read only assemblies. The quality of our assembly is also benchmarked against other bivalves genome assemblies and we present an initial phylogenomic analysis for the class Bivalvia, which illustrates the value and potential of the increasing number of high quality genomic data sets for phylogenetics.

  doi: 10.3389/fgene.2019.01158