Synbio Technologies’ immune repertoire sequencing and bioinformatics analysis:
1. Library Construction
Synbio Technologies performed 5’ RACE reverse transcription of the extracted RNA and constructed the library.
2. Library Quality Control
The Qubit Fluorometer was used to perform preliminary quantification after library construction. Following dilution, insert size was determined based on the Agilent bioanalyzer. Once expected requirements were met, the effective concentration of the library (>2nM) was then accurately quantified by qPCR to ensure library quality.
3. Library Sequencing
Different libraries were pooled and sequenced by Illumina Miseq/HiSeq according to the effective concentration and the required amount of target data. Four fluorescent labeled dNTPs, DNA polymerase, and adaptors were added to the flow cell for the amplification. When each sequence cluster extended and generated the complementary chain, the corresponding fluorescent signals were released and captured after each new dNTP was added. The optical signals were then transformed into sequencing peaks by professional software to obtain the sequence information of the fragments.
4. Data Analysis
A quality assessment of the sequencing data was conducted.
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Per Base Sequence Quality
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Per Sequence Quality Scores
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Per Base Sequence Content
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Per Base GC Content
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Per Sequence GC Content
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Per Base N Content
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Sequence Length Distribution Reads
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Duplicate Sequences
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DNA Synthesis
Vector Selection
Molecular Biology
Oligo Synthesis
RNA Synthesis
Variant Libraries
CRISPR Libraries
Oligo Pools
Virus Packaging
Gene Editing
Protein Expression
Antibody Services
Peptide Services
DNA Data Storage
Standard Oligo
Standard CRISPR Libraries
Standard CRISPR Plasmid
ProXpress
Protein Products
