DNA Sequencing

Sample Preparation Instructions

Pricing

  • We guarantee competitive prices for both Non-profit and private sector organizations.
  • Please call in for a quote.

Benefits

  • The most flexible sequencing service with the highest success rate
  • Skilled interpretation of your sequencing results
  • Trouble-shooting based on our expertise and our proposals to deal with sequencing difficulties (hairpin, GC-rich templates)
  • Additional services (template amplification, PCR purification) are included in the service package
  • Optional sequence editings and sequence alignment are available free of charge

Quality

  • Average reading lengths: 700-1000 bases.
  • Color printouts of chromatographs are provided free of charge upon request.
  • Optional sequence alignment saves time when analyzing sequencing results.
  • Sequence editings are provided upon request. Sequence editing means a manual proofread performed by an experienced staff to correct mistakes /uncertainities in sequences generated by the automatic base callor.
  • Trouble shooting: We propose further trouble shooting steps based on the sequencing results. However, it is your decision whether we continue or stop the sequencing job.
  • Repeat policy: Failed samples will be repeated based on our interpretation of your sequencing results, and the repeat is free of charge.

Turnaround

The general turnaround time is 24 hours. Add another 24 hours for E.coli strains.

Colony PCR

Pricing

  • We guarantee competitive prices for both Non-profit and private sector organizations.
  • Please call in for a quote.

This protocol is designed to quickly screen for plasmid inserts directly from E. coli colonies. The plasmid should be high copy number such as pUC18 pUC 19, or pBluescript, etc. Even though blue/white screening can be used to determine if inserts are present, this technique can be used to determine insert size and/or orientation in the vector. We have an optimized colony PCR protocol to tailor your insert size and plasmid type. After colony PCR is done, a small amount of product will be checked by by agarose gel electrophoresis. If predicted band is observed, ExoSAP process will be carried out to remove excess primer and dNTP, and then DNA sequencing will be executed.

Template Amplification (RCA)

Pricing

  • We guarantee competitive prices for both Non-profit and private sector organizations.
  • Please call in for a quote.

Template amplification uses bacteriophage Phi29 DNA polymerase to in vitro amplify plasmid DNA, also called rolling circle amplification (RCA). RCA has a strong preference of circular DNA templates like plasmid DNA. Plasmid amplified by RCA is suitable for DNA sequencing. You can submit DNA samples in case you have limited DNA samples, or E.coli strains. Turnaround time for RCA amplification is 24 hours. When combined with DNA sequencing, turnaround time for the whole process is 48 hours.

Nucleic Acid Extraction

Pricing

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Plasmid preparation

A plasmid preparation is a method used to extract and purify plasmid DNA. We used Qiagen Miniprep to Maxiprep kits to purify plasmid DNA for your need. We accept various forms of samples, such as bacteria culture, bacteria colony plate, bacteria seed or ligation product. After plasmid preparation is done, DNA sequencing will be preformed. Plasmid DNA or bacteria seed can be returned to you upon request.

Genomic DNA preparation from tissue

A customized protocol is available for extracting genomic DNA from animal tissue (commonly mouse tail). This yields DNA suitable most downstream applications, such as genotyping, Southern blotting, sequencing, etc. The genomic DNA can be returned to you upon request.

Genotyping

Pricing

  • We guarantee competitive prices for both Non-profit and private sector organizations.
  • Please call in for a quote.

Genotyping refers to the process of determining the genotype of an individual by the use of biological assays. Our methods of doing this include PCR, DNA sequencing and qPCR. The technology is important in clinical research for the investigation of disease-associated genes.

Genotyping applies to a broad range of research interests including SNP detection, transgenic/knockout determination and strain determination.

Cosmid and BAC Sequencing

Reactions

Sequencing of cosmid and Bac-DNA is similar to sequencing of plasmid templates except that more DNA is needed. 2 ug Cosmid/Bac DNA is needed for direct sequencing. There is no additional charge for Cosmid/Bac sequencing.

We also offer the whole cosmid- and Bac-DNA shotgun sequencing. Please inquire about details.

High Throughput Sequencing Services

Pricing

  • We guarantee competitive prices for both Non-profit and private sector organizations.
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Reactions

Single-pass sequencing reactions are performed in 96-well PCR plates.

Sample Submission

  • The DNA samples and the primers should be pre-mixed in defined volume.
  • Samples must be submitted in 96-well PCR plate format to preserve samples' internal order.
  • Common primers can be added by our staff free of charge upon request.

Plasmids

We also accept E.coli strains instead of purified plasmid DNA. We in vitro amplify plasmids from bacteria for DNA sequencing.

PCR-products

We purify PCR products from primers and dNTP using Exonuclease and gel filtration.

PCR Purification

Pricing

  • We guarantee competitive prices for both Non-profit and private sector organizations.
  • Please call in for a quote.

We purify PCR products from primer and dNTP by using exonuclease and gel filtration columns.

PCR products should have a single distrinct band on agarose gel, otherwise, gel purification should be pursued.