• DNA Sequencing
• Ready-to-Run
• Sequencing from purified templates
• Full service sequencing
• Genotyping
• Quantitative and real time PCR
  • Ready-to-Run Taqman
  • Full services Taqman
• Bacterial and Fungal Identification by rDNA Sequencing Analysis
  • Bacterial Identification 500
  • Bacterial Identification Full Length
  • Fungal Identification 600
• Transgenic Mouse Genotyping

---

• DNA Sequencing:

  • Ready-to-Run Samples:

    Ready-to-Run Samples: Laragen provides sequencing from "Ready-to-Run" samples. With this service, customers will perform their own sequencing reactions in 8 strip PCR tubes in 10 ml of reaction volume.   We provide FREE Bigdye cleaning for ready-to-run samples.   The turnaround time is overnight.

    Please following the following protocol and parameters to set up ready-to-run sequencing reaction. Most of ready-to-run sequencing problems are caused by too much or too little DNA.

    Ready-To-Run Sequencing Reaction Setup Protocol

    Cycle Sequencing Run Parameter (based on ABI 9700 Thermocycler)

  • Sequencing from purified templates:

    Sequencing from Purified Templates: We provide sequencing from virtually all types of templates; plasmids, PCR products, lambda phage, BACs, PACs, etc. We provide 10 common primers free of charge, however we require an aliquot of custom primers with your templates. Turnaround time is next day.

    Sample requirements: We require all plasmids to be purified with commercial columns. We recommend Qiagen miniprep kit. We do not recommend gel extraction for PCR products if the PCR products have single bands. . The following table outlines the required concentrations for different types of templates for successful sequencing.

     

    Template	Concentration(ng/ul)	Volume(ul)
    Plasmids(up to 15Kb)	~100	10 - 20
    PCR products	~25	10- 20
    BACs, PACs (ends only)	~200	25

    Primer requirements: We will provide T7, T3, T7 terminator, SP6, M13 Forward,  M13 Reverse, RV3,  BGHR, pGEX5' and pGEX3' primers. The general rules for designing sequencing primers are the same as for PCR primers. The primer does not need special purification. The work solution of primer should be at 10 uM (10 pmole/ul). We use 5 pmole of primer per DNA sequencing reaction.

  •   Full service sequencing

    Full service sequencing: Full service sequencing is available from bacterial cultures. The clients can provide us with either 96 well growth blocks or individual 3-5ml cultures. We return the sequences and Qiagen purified plasmids DNA with the concentration determined by fluorescent measurement upon request.

    Sample requirements: Cultures should be grown in LB in 96-well growth blocks or any 16 or 17 X 100 mm culture tubes. Growth blocks require 22 to 24 hours of incubation with 250 rpm shaking. Samples should be centrifuged, decanted and the pellets should be sent to us frozen.

    We also prepare DNA templates with Templiphi from GE Health Sciences,  especially with large volume of samples.  Please contact us for the culture condition for Templiphi.

• Genotyping and DNA Fragment Analysis:

  Laragen is pleased to offer genotyping services and DNA fragment analysis. Our ABI 3730 sequencers and ABI 7900HT Fast Realtime PCR System provide us different platforms for DNA fragment analysis and SNP genotyping.  Please contact us to discuss your specific requirements.

• Quantitative and Real Time PCR

  • Ready-to-Run Taqman: Investigator design and synthesize the primer/probe sets. They also set up the PCR plates. We run the plates in an ABI 7700 Taqman Real Time Sequence Detection system and assist in analysis of the data.

  • Full services Taqman:  Investigators provide RNA or DNA samples. Laragen designs and synthesizes the primer and probe sets, set up the PCR reactions, optimizes PCR conditions and provides analyzed results.

• Bacteria and Fungi Identification by rDNA Sequencing Analysis

  Based on the length of rDNA used, Laragen currently provides two tests of bacterial identification by 16S rDNA and one test of fungal identification by 28S rDNA. We use Genebank databases to identify the unknown bacteria and fungi. The advantages of bacterial and fungal identification by rDNA sequences are fast turnaround time and accurate results even for problematic bacteria and fungi. All reports include  the consensus sequence of the tested bacterial/fungal sequences, the alignment of top 10 matches at species or genus level and a phylogenetic tree with these top 10 sequences. Our turnaround time is 5 business days.

  • Bacterial Identification 500
    • Extract genomic DNA from bacterial colony or culture.
    • PCR amplify and sequence the first 500 bp of 16S rDNA
    • Identify the unknown bacterium using GenBank Databases
    • Align the unknown sequence with top 10 matches at species or genus level and a phylogenetic tree is constructed using neighbor-joining algorithm.
  • Bacterial Identification Full Length
    • Extract genomic DNA from bacterial colony or culture.
    • PCR amplify and sequence the full length of 16S rDNA
    • Identify the unknown bacterium using GenBank Databases
    • Align the unknown sequence with top 10 matches at species or genus level and a phylogenetic tree is constructed using neighbor-joining algorithm.
  • Fungal Identification D1D2
    • Extract genomic DNA from fungal colony or culture.
    • PCR amplify and sequence the first 600 bp of 28S rDNA (D1/D2 region)
    • Identify the unknown fungus using GenBank Databases
    • Align the unknown sequence with top 10 matches at species or genus level and a phylogenetic tree is constructed using neighbor-joining algorithm.

    You can download Bacterial and Fungal ID Service Request Form.

  • Transgenic mouse genotyping

    Laragen provides transgenic, knockout, knockin and shRNA transgenic mouse genotyping services since 2002. Most of mouse genotyping services are performed with probe based realtime PCR. Our goal is to provide fast and accurate mouse genotyping services to researchers from both academic and industrial institutions. Our turnaround time is 24 to 48 hours. Same day rush services are also available upon request. Our mouse genotyping accuracy rate is above 99.95%. We accept a variety of mouse tissues samples that include tails, ears, toes, brains and other tissues. We also accept genomic DNA samples. Depending on the number of samples and nature of the tests, we either waive the initial set-up fee or only charge a nominal set-up fee to cover the reagent cost. Four types of mouse genotyping services are available at Laragen:

    • Probe based Realtime PCR
    • Gel based PCR
    • Transgene copy number based on quantitative PCR
    • Trinucleotide repeat number analysis

    You can download Laragen Mouse Genotyping Service Request Form.

    We provide both volume discount and multiple alleles discount. Please contact us to inquire price information.