CD Genomics provides a fast, one-stop bacterial RNA sequencing solution from the quality control of sample to comprehensive data analysis. Please contact us for more information and a detailed quote.
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Bacterial rna sequencing
1. Bacterial RNA Sequencing
Besides eukaryotic RNA sequencing service, CD Genomics is also
dedicated to providing prokaryotic RNA sequencing service to advance
your bacterial gene expression profiling needs by utilizing the latest
techniques.
The Introduction of Bacterial RNA Sequencing
In recent years, high throughput sequencing of cDNA libraries (RNA-
Seq) has emerged as a powerful technology for profiling gene
expression, discovering previously unannotated genes, and mapping
transcriptome architecture in a wide variety of bacterial species. With
the application of RNA-seq derivative approaches, we can gain
biological insights into the bacterial world and aspire to uncover the
mysteries involving gene expression, organization and other functional
genomic features. Bacterial transcriptome
and metatranscriptome information is important for predicting resistance
to specific antibiotics, understanding host-pathogen immune
interactions, quantifying gene expression changes, and tracking disease
progression.
There are obvious differences of cDNA library construction between
bacterial RNA-Seq and Eukaryotic RNA-Seq. Bacterial RNA-Seq
workflow’s first step is selection of mRNA transcripts. Ribo-Zero
ribosomal RNA reduction chemistry is used in place a poly-A tail
selection which makes this method particularly suited for bacteria
mRNA lack a poly-A tail. Following purification, the mRNA is fragmented
into small pieces, and copied into first strand cDNA using reverse
transcriptase and random primers. Strand specificity is achieved by
replacing dTTP with dUTP in the Second Strand Marking Mix (SMM),
followed by second strand cDNA synthesis. Through the use of strand-
specific RNA-Seq, a more complete understanding of the transcriptome
could be achieved, this has the potential to identify new levels of
regulation of gene expression.
Project Workflow
Sample Requirements:
2. 1. RNA amount: Total RNA≥3 ug (without degradation or DNA
contamination)
2. RNA purity: OD260/280 = 1.8~2.2; OD260/230 ≥1.8
3. RNA quality: 28S:18S≥1.5,RIN≥7
Sequencing Strategy:
HiSeq X ten, 150 PE, 1~3 G/per sample
Data Analysis
Transcriptome with no Reference
Transcriptome Assembly
Transcriptome/Unigene Length Statistic
Transcriptome Function annotation, including NR, NT, Gene Ontoloy,
KEGG pathway, KOG, Swissport, Protein Family
Differential expression analysis
CDS Predictive Parsing
SNP/InDel Analysis
… (more upon request)
Transcriptome with Reference
Transcriptome Assembly
Transcriptome/Unigene Length Statistic
Transcriptome Function annotation, including NR, NT, Gene Ontoloy,
KEGG pathway, KOG, Swissport, Protein Family
Differential expression analysis
CDS Predictive Parsing
SNP/InDel Analysis
… (more upon request)
Transcriptome with Reference
Reference Genome mapping
Transcriptome Function annotation, including NR, NT, Gene Ontoloy,
KEGG pathway, KOG, Swissport, Protein Family
Differential expression analysis
Alternative splicing
Antisense Transcript Analysis
SNP/InDel Analysis
… (more upon request)
CD Genomics provides a fast, one-stop bacterial RNA sequencing
solution from the quality control of sample to comprehensive data
analysis. Please contact us for more information and a detailed quote.