Provide accuracy of 100% per purification for large scale genome projects to FDA submission projects
For Use With (Application)
PCR product gel purification service
Ranging from large-scale genome projects to FDA submission projects with a guaranteed accuracy of 100%.
DNA Sequencing Services Single Pass Sequencing Services
A variety of templates may be submitted for sequencing including plasmids, PCR products, and single strand templates
A reaction is considered to be one template sequenced with one primer
DNA can be prepared if an overnight culture or plate is provided.
Purified Plasmid DNA should be submitted at the optimal concentration of 0.2μg/μL, and 1–2μg of DNA should be provided per sequencing reaction.
PCR products should be gel or column purified.
Provide 10–20μL of purified PCR products at a concentration of 0.02–0.05μg/μL
PCR products can be gel or column purified if 50μL are provided
Results are returned as data file that includes a chromatogram and text file
Primer Walking Sequencing
The method of choice for sequencing DNA fragments between 1.2 and 7kb
Primers will be designed and synthesized as needed to obtain single (confirmation quality) or double strand (publication quality) contiguous sequence information for the region of interest
For double-strand sequencing, provide 10μg/kb
For single-strand sequencing, provide 5μg/kb
Results include a chromatogram, text file, and consensus sequencing
FDA Submission Sequencing
Sequence determination with a 100% accuracy level
Rigorous quality control and documentation are provided and data meets compliancy level suitable for submission to the FDA or other regulatory agencies
A report with consensus sequencing and contig alignment map will be returned
Shotgun Sequencing Services
Shotgun libraries can be created from Cosmids, BACs, PACs, or genomes
Typically, insert sizes range from 1–3kb and are cloned into pUC18
Full length sequencing services provide a guaranteed double-strand sequence with an accuracy of 99.99%
Provide 100μg of purified DNA or a clone from which to grow and purify DNA and indicate the number of clones needed to complete the project
The resultant pUC18 shotgun library can be shipped on agar plates
Note: contamination of plasmid or BAC DNA by genomic DNA will make sequencing of the library very difficult and can necessitate reconstruction of the library
Template Preparations Any template can be prepared for sequencing or library construction, using proprietary protocols to ensure optimal purity and yield. All samples remain the property of the customer and can be returned upon the completion of the project. Template Miniprep and Midiprep.
DNA can be prepared and purified if an overnight culture or plate is provided
Minipreps produce a yield of approximately 10μg for most high-copy plasmids
Midipreps produce a yield of approximately 75–100μg for most high-copy plasmids
Template BAC, PAC, Cosmid, or P1 Prep
DNA can be prepared or purified from a provided plate or overnight culture
The DNA is purified by CsCl gradient centrifugation (2×) to remove host genomic DNA
Yield is approximately 25μg of DNA in water or TE
Standard unmodified custom oligonucleotides can be synthesized from a submitted sequence
Alternately, oligonucleotides may be provided as starting material at a concentration of 3.2μL with a total volume of no less than 20μL for single tube reactions or 200μL for samples in 96-well format
The synthesized oligos may be returned to the customer or used for subsequent sequencing