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dna methylation sequencing

Specialty DNA Sequencing

DNA Methylation Sequencing

dna methylation sequencing graph Methylation of cytosine within regulatory genetic elements is a central mechanism in epigenetic regulation of gene expression. Our DNA methylation sequencing services will map methylation sensitive areas of genes.

We offer a full service analysis to determine the location and extent of cytosine methylation patterns in the DNA. Genomic DNA is purified and reacted with a bisulfite reagent that converts cytosine to uracil, but leaves 5-methylcytosine intact. A target region from unreacted- and bisulfite-treated DNA is PCR amplified. During amplification, uracil is treated as a thyimidine and 5-methylcytosine as a cytosine. DNA sequencing of the PCR products from the unreacted and bisulfite-treated DNA reveals the location of methylated cytosines.

When the methylation pattern of a DNA fragment is compared across treatments, (i.e., age, exposure to mutagenic chemicals, metastasis) this analysis is able to pinpoint putative epigenetic elements in the DNA. Variations of our services include high throughput screening of new and previously defined methylation-sensitive DNA regions across treatment conditions.

ACGT, Inc. offers a methylation analysis package including:

  • DNA extraction
  • Bisulfite conversion of DNA
  • Design and optimization of primers for PCR amplification and DNA sequencing
  • PCR amplification of target DNA
  • Double strand DNA sequencing of unreacted and bisulfite-treated DNA
  • Comparison of methylation patterns across treatment conditions

Genomic DNA Re-Sequencing

Genotyping by DNA sequencing provides insight into mutations at a single nucleotide level.  We provide full genotyping DNA re-sequencing services that include assay design and validation with primer design and optimization, genomic DNA extraction, PCR amplification, double-strand DNA sequencing, DNA sequencing data editing and alignment, and identification of single nucleotide polymorphisms (SNPs), deletions, insertions and inversions.  Pre-validated assays for human genes are used when available.  This service is also available under GLP-compliant conditions.

Additional Re-Sequencing Services:

  • Use of pre-validated assay sets for human genes
  • Pre-validated assay sets for human genes
  • Large sequencing capacity for short turnaround time
  • Optional follow-up validation services with Taqman SNPs Assay and Fragment Analysis

BAC End Sequencing

DNA sequencing of BAC and COSMID DNA with gene-specific or vector-specific primers is available including full gene DNA sequence analysis. Our services include colony picking, bacterial cell growth, DNA extraction from clones, DNA sequencing and Data Analysis including sequence alignment and comparative analysis from a single sample to a 96-well platform for high-throughput scale processing.

Microorganism Sequencing

Bacterial Identification By DNA Sequencing:

fungal_id_sequencingOur bacterial identification services are based on the phylogenetic analysis of 16S rRNA gene by DNA sequencing. The technique can be applied to monitor environmental cleanness of a production area, monitor fermentation processes, or research in a clinical lab, or analysis of novel isolates. We offer services with GLP compliant or without depending on your application.

Service Description:

  • Purification of genomic DNA
  • PCR amplification of 16S rRNA gene
  • Double strand sequencing
  • Identification to the genus or species level by comparing sequence data to PE Biosystems' MicroSeq® Database and GenBank public database
  • Analysis report including top ten matches and phylogenetic trees, electropherograms and consensus sequence
  • Three to five-day turnaround time

We offer two different levels of services: Service #1 and #2. Bac ID #1 analyzes first 500 bases of the gene while Bac ID #2 includes full-length sequence of 16S rRNA gene. fungal_id_sequencing

Fungal Identification By DNA Sequencing:

  • Growth of the fungus on solid medium if necessary
  • Extraction of genomic DNA
  • PCR amplification of the 300-500 base pair sequence (Our fungal identification services are based on the phylogenetic analysis of the D2 region of the large-subunit (LSU) rRNA gene)
  • Double strand DNA sequencing
  • Identification at the genus or species level by comparing sequence data to GenBank public database
  • Analysis report including top ten matches and phylogenetic trees, electropherograms and consensus sequence
  • Three to five-day turnaround time

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