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   Next-Generation DNA Sequencing (NGS)


To request a quotation or for placing orders, please download our order form (DNA Order form or RNA Order Form) and email it to sequencing@biobasic.com. Our NGS Specialist will get back to you within 24 hours (during a business day; EST) to review project details and discuss pricing.

Features:

  • Comprehensive Sequencing platforms- Bio Basic has 454, Illumina, Ion Torrent sequencing and Sanger sequencing platforms. We could provide optimal solutions with flexible use of these platforms according to the research purpose or the requirement of experiments.
  • Cost-effective - We understand budgets are tight and thus will ensure the most competitive rates for projects.
  • Powerful Bioinformatics analysis and follow-up Services - We provide customized bioinformatics analysis services and reports, to assist with addressing your research problems, data visualization queries and concerns.

Overview of Next Generation Sequencing (NGS):

Next Generation Sequencing (NGS) is a fairly new technology that has rapidly expanded the capabilities of molecular biology and genomics. It goes beyond the capabilities of traditional DNA sequencing and allows the sequencing of whole genomes at a fraction of the time and cost. Although NGS workflow involves quite diverse techniques and biochemistry at each step from template library preparation, fragment amplification, to sequencing, they all adopt a massive matrix configuration popularized by microarray analysis: A vast number of DNA samples on the array are simultaneously analyzed in parallel. First, massive parallelism can be achieved through ordered or disordered array configuration that offers high degree of information density. This dramatically increases the overall throughput of the sequencing operation. Second, no electrophoresis steps are required. So it's widely used in Systems Biology, Function Genomics, Epigenomics, Exon Sequencing, Transcriptome, etc.

Our Next NGS Services include:

1. Microbiome sequencing

Microbiome sequencing allows the determination of the marker gene sequence of bacteria, archaea or fungi such as 16s rRNA,18s rRNA or ITS .The microbial diversity, population structure, evolutionary relationship, functional activity, mutual cooperation and the relationship between the microbial diversity and the environment are studied by using marker gene screening and comprehensive sequencing analysis.

Project workflow:

MiSeq platform:


Genomic DNA QC

16S / 18S / ITS region amplification

Quantitative mixing of amplified products

MiSeq sequencing

Data output

Bioinformatics analysis

Final project report

 

Ion Torrent platform:


Genomic DNA QC

16S / 18S / ITS region amplification

Quantitative mixing of amplified products

Emulsion PCR amplification

Ion Torrent sequencing

Data output

Bioinformatics analysis

Final project report

 


Project Stage

Content

Turnaround

Target region amplification

gPCR and library construction

1 to 5 days

High throughput sequencing

MiSeq platform

20 days

Data analysis

Comprehensive Data Analysis and Visualization

20 days

Note: After DNA samples are extracted, the regions are selectively amplified (such as the corresponding regions of bacteria 16S V1-V9). The amplified products are reconstructed and sequenced, followed by bioinformatics analysis. Bioinformatics analysis includes OTU generation, sampling adequacy analysis, abundance and diversity analysis, inter-flora analysis, hypothesis validation analysis, phylogenetic tree analysis, and more.

 

Required, for project initiation:
Samples:
Total DNA or packaged raw samples (such as soil, faeces, water and mouth) should be provided with their names clearly labelled. Samples should be shipped via refrigerated transport. For more info. on specific requirements by sample types, see "NGS sample requirements".
Information:
Provide information on the species as well as the control and experimental samples.

Deliverable (Report) contains:
Sequencing Raw Data
Experimental Results Class File
Analysis Report and Analysis Results File.

 

2. Small Genome sequencing

With the rapid development of bacterial genome research, genome sequencing has gradually become essential for basic microbial research. The new generation of high-throughput sequencing significantly reduces the cost and time of genome sequencing, thereby making it more accessible for researchers to carry out microbial genome sequencing projects. Entire genomes of multiple bacteria, viruses, or other microbes can be sequenced simultaneously using this method of high-throughput sequencing.

Project workflow:

HiSeq platform:


Genomic DNA Extraction

Genome 5K MP library construction

HiSeq sequencing

Data output

Bioinformatics analysis

Data output

 

MiSeq platform:


Genomic DNA Extraction

Genome PE library construction

MiSeq PE sequencing

Data output

Bioinformatics analysis

Data output

 


Project Stage

Content

Turnaround

Genomic DNA extraction

Bacterial genome extraction

1 to 3 days

Bacteria sequencing 16S full
length verification
 
1 to 3 days

Library construction

PE library construction

2-5 days

High throughput sequencing

MiSeq, HiSeq platform,
sequencing 2G data

20 days

Data analysis

Comprehensive Data Analysis and Visualization

20 days

 

Required, for project initiation:
Samples:
Human tissue or total genomic DNA samples should be provided with their names clearly labelled. Samples should be shipped via refrigerated transport. For more info. on specific requirements by sample types, see "NGS sample requirements".
Information:
Provide information on the species as well as the control and experimental samples.

Deliverable (Report) contains:
Sequencing Raw Data.
Experimental Results Class File.
Analysis Report and Analysis Results File.

 

3. Exome sequencing

Exome sequencing (aka whole exome sequencing, WES) is an analysis method for sequencing all the protein coding regions of all genes.

Sequencing of exon subgroups can be used to find single gene diseases, complex diseases (such as diabetes, obesity and other metabolic syndromes) and even cancer genes or genetic susceptibility. Because of this, exome sequencing shows promising clinical applications as the methodology can be used to study rare genetic diseases by sequencing only exons in a genome.

Project workflow:


Genomic DNA QC

Genomic library construction

Exon region capture

Bridged PCR single molecule amplification

HiSeq sequencing

Data output

Bioinformatics analysis

Final project report

 


Project Stage

Content

Turnaround

Exon region capture

Whole exon capture by Aglient Sureselect kit

5 days

High throughput sequencing

HiSeq platform, sequencing
10G (optional 8G) data volume

20 days

Data analysis

Comprehensive Data Analysis and Visualization

20 days

 

Required, for project initiation:
Samples:
Human tissue or total genomic DNA samples should be provided with their names clearly labelled. Samples should be shipped via refrigerated transport. For more info. on specific requirements by sample types, see "NGS sample requirements".
Information:
Provide information on the species as well as the control and experimental samples.

Deliverable (Report) contains:
Sequencing Raw Data.
Experimental Results Class File.
Analysis Report and Analysis Results File.

 

4. ChIP-sequencing (ChIP-seq)

The ChIP-seq technology combines chromatin immunoprecipitation with high throughput sequencing. It is a powerful tool for studying the interaction between proteins and DNA in vivo and can also be used to study the relationship between transcription factors and gene expression.

Through high-throughput sequencing, the binding distribution of the target protein across the whole genome can be obtained at a time, and the binding site of the target protein and the binding motif can be obtained.

The millions of sequence tags obtained are accurately mapped to the genome to obtain DNA segment information that interacts with histones, transcription factors, etc. within the whole genome.

Project workflow:


ChIP DNA QC

ChIP DNA library construction

PCR Library enrichment

Bridged PCR single molecule amplification

HiSeq sequencing

Data output

Bioinformatics analysis

Final project report

 


Project Stage

Content

Turnaround

Library construction

Library construction

1 to 5 days

High throughput sequencing

HiSeq platform, sequencing
10 Million reads

20 days

Data analysis

Comprehensive Data Analysis and Visualization

20 days

 

Required, for project initiation:
Samples:
Final DNA from ChIP experiment should be provided with their names clearly labelled. Samples should be shipped via refrigerated transport. The ChIP project requires at least 50 ng of DNA, subject to Qubit 2.0 testing. For more info. on specific requirements by sample types, see "NGS sample requirements".
Information:
Provide information on the species as well as the control and experimental samples.

Deliverable (Report) contains:
Sequencing Raw Data.
Experimental Results Class File.
Analysis Report and Analysis Results File.

 

5. RNA Sequencing (RNA-Seq)

RNA Sequencing is a transcriptome study based on a new generation of sequencing techniques. RNAs of biological samples are extracted. Then, mRNA is enriched in the sample, followed by reverse transcription into cDNA for high-throughput sequencing. On the basis of the fragment assembly or reads mapping to reference genome, Goodunderstanding of the gene expression from the biological samples can be achieved Changes in gene expression levels for key genes can also be analysed.

Project workflow:


Total RNA QC

mRNA purification and fragmentation

mRNA Library construction

Bridged PCR single molecule amplification

HiSeq sequencing

Data output

Bioinformatics analysis

Final project report

 


Project Stage

Content

Turnaround

RNA extraction

Animal/ Plant RNA extraction

1 to 3 days

RNA library

Transcriptome RNA library construction

3 to 5 days

High throughput sequencing

HiSeq platform, sequencing
6G data volume

20 days

Data analysis

Comprehensive data analysis and visualization

20 days

 

Required, for project initiation:
Samples:
Tissue or total RNA samples should be provided with their names clearly labelled. Samples should be shipped with dry ice. For specific requirements by sample types, see "NGS sample requirements".
Information:
Provide information on the species as well as the control and experimental samples.

Deliverable (Report) contains:
Sequencing Raw Data.
Experimental Results Class File.
Analysis Report and Analysis Results File.

 

6. Small RNA sequencing

Small RNA, including microRNA, ncRNA, siRNA, snoRNA, piRNA and rasiRNA acts as an important regulatory factor in biological processes. Small RNA high-throughput sequencing can quickly and comprehensively detect small RNAs within a given sample.

The process involves RNA sample extraction and purification followed by PAGE gel electrophoresis to isolate the target small RNA. The cDNA Library is then constructed for the sample to undergo high throughput sequencing and data analysis.

Project workflow:


RNA QC

Connecting ends

Reverse transcription & PCR of small RNA library

Bridged PCR single molecule amplification

HiSeq sequencing

Data output

Bioinformatics analysis

Final project report

 


Project Stage

Content

Turnaround

RNA extraction

Small RNA extraction

1 to 3 days

RNA library

Library construction

3 to 5 days

High throughput sequencing

HiSeq platform, sequencing
10 Million reads

20 days

Data analysis

Comprehensive Data Analysis & visualization

20 days

 

Required, for project initiation:
Samples:
Tissue or total RNA samples should be provided with their names clearly labelled. Samples should be shipped with dry ice. For more info. on specific requirements by sample types, see "NGS sample requirements".
Information:
Provide information on the species as well as the control and experimental samples.

Deliverable (Report) contains:
Sequencing Raw Data.
Experimental Results Class File.
Analysis Report and Analysis Results File.