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Prokaryotic mRNA sequencing

mRNA sequencing empowers the comprehensive profiling of all mRNA transcripts within cells under specific conditions. This cutting-edge technology serves as a potent tool, unveiling intricate gene expression profiles, gene structures, and molecular mechanisms associated with diverse biological processes. Widely adopted in fundamental research, clinical diagnostics, and drug development, mRNA sequencing offers insights into the intricacies of cellular dynamics and genetic regulation. Our prokaryotic mRNA sample processing is tailored for prokaryotic transcriptomes, involving rRNA depletion and directional library preparation.

Platform: Illumina NovaSeq X


Service Details

Bioinformatics

Demo Results

Featured Publications

Features

● RNA sample processing involved rRNA depletion followed by directional RNA library preparation.

● Bioinformatic analysis based on alignment to a reference genome

● Analysis includes gene expression and DEGs but also transcript structure and sRNA analysis

 

Advantages

● Rigorous Quality Control: we implement core control points across all stages, from sample and library preparation to sequencing and bioinformatics. This meticulous monitoring ensures the delivery of consistently high-quality results.

● Strand-specific sequencing data: due to the RNA library preparation being directional, enabling identification of anti-sense transcripts.

● Complete analysis tailored to prokaryotic transcriptomes: the bioinformatic pipeline includes not only analysis of the gene expression but also analysis of the transcript structure, including identification of operons, UTRs and promoters. It also includes analysis of sRNAs, namely annotation and prediction of secondary structure and targets.

● Post-Sales Support: our commitment extends beyond project completion with a 3-month after-sale service period. During this time, we offer project follow-up, troubleshooting assistance, and Q&A sessions to address any queries related to the results.

Sample Requirements and Delivery

Library

Sequencing strategy

Data recommended

Quality Control

Poly A enriched

Illumina PE150

1-2 Gb

Q30≥85%

Sample Requirements:

Conc.(ng/μl)

Amount (μg)

Purity

Integrity

≥ 50

≥ 1

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Limited or no protein or DNA contamination shown on gel.

RIN≥6.5

 5.0≥28S/18S≥1.0;

limited or no baseline elevation

 

Recommended Sample Delivery

Container: 2 ml centrifuge tube (Tin foil is not recommended)

Sample labeling: Group+replicate e.g. A1, A2, A3; B1, B2, B3... ...

Shipment:

1. Dry-ice: Samples need to be packed in bags and buried in dry-ice.

2. RNAstable tubes: RNA samples can be dried in RNA stabilization tube(e.g. RNAstable®) and shipped in room temperature.

 

Service Work Flow

sample delivery

Sample delivery

Library Preparation

Library construction

Sequencing

Sequencing

Data analysis

Data analysis

After sale Services

After-sale services


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  • Bioinformatic Analysis Workflow

    ProkaryoticmRNA-01

    Includes the following analysis:

    ● Raw data quality control

    ● Alignment to reference genome

    ● Library quality assessment: RNA fragmentation randomness, insert size, and sequencing saturation

    ● Functional annotation of predicted coding genes

    ● Expression analysis: correlation and Principal Component Analysis (PCA)

    ● Differential Gene Expression (DEGs)

    ● Functional annotation and enrichment of DEGs

    ● sRNA analysis: prediction, annotation, target, and secondary structure prediction

    ● Transcript structure analysis: operons, starting and ending positions, Unstranslated region (UTS), promoter and SNP/InDel analysis

    Sequencing saturation

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    Functional annotation of coding genes

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     Correlation between samples

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    Differential Expressed Genes (DEGs) analysis

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    Functional enrichment analysis

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    sRNA annotation

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    Explore the advancements facilitated by BMKGene’s Nanopore full-length mRNA sequencing services in this featured publication.

     

     Guan, C. P. et al. (2018) ‘Global Transcriptome Changes of Biofilm-Forming Staphylococcus epidermidis Responding to Total Alkaloids of Sophorea alopecuroides’, Polish Journal of Microbiology, 67(2), p. 223. doi: 10.21307/PJM-2018-024.

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