† The PCR process is covered by patents owned by Roche Molecular Systems, Inc.,
and F. Hoffman-La Roche, Ltd.
* All trademarks are property of their respective owners.
Beckman Coulter, the stylized logo, Agencourt, CleanSEQ and SPRI are registered trademarks of Beckman Coulter, Inc.
Single Pass Sequencing
Beckman Coulter Genomics Single Pass service is highly optimized for high-throughput sequencing of plasmids or PCR† amplicons submitted in 96 or 384 well formats. Fueled by a high capacity, high performance Sanger sequencing pipeline Beckman Coulter Genomics Single Pass sequencing service is designed for:
- Rapid project turnaround
- High sequencing pass rates
- Long Phred 20 read lengths
Samples and Sequencing Primers
Samples
Beckman Coulter Genomics accepts 96 and 384 well plates of the following for Single Pass sequencing:
- Glycerol stocks
- Transformed libraries
- Agar plated libraries
- Unpurified PCR samples
Please see Beckman Coulter Genomics Sample Submission Guidelines for submission tips that optimize Single Pass project turnaround times and enable the highest level of DNA sequencing quality.
Sequencing Primers
Single Pass sequencing of plasmids or tailed PCR products can be performed using universal primers supplied by Beckman Coulter Genomics. Alternatively, customers may submit custom sequencing primers arrayed in a 96 or 384 well plate format corresponding to their associated plate of templates.
Universal Primers for available for Single Pass sequencing
- Universal T7
- M13-FWD
- M13-REV
- SP6
- Universal T3
- KBR/TJ
- 2BP Clamp
- SeqL-A
- SeqL-B
- T7 Terminator
- CMV Forward
- BGH Reverse
Purification and Sequencing
Experience has proven that coupling automated SPRI template purification with Agencourt CleanSEQ dye terminator removal from sequencing reactions facilitates the generation of long, high quality Sanger sequencing reads. Upon receipt all Single Pass projects therefore proceed through the following proven process:
- Purification of plasmids and PCR amplicons using SPRI technology
- DNA sequencing using BigDye* Terminator v3.1
- Post reaction dye terminator removal using Agencourt CleanSEQ
- Sequence delineation on an ABI PRISM 3730xl
- Base calling and data compilation
Results
DNA sequencing results from the Single Pass pipeline are delivered via a secure FTP site. Data delivery is overseen by a Beckman Coulter Genomics Project Manager to ensure simple and timely transfer. Compiled data is available in multiple file formats including:
- FASTA
- .AB1
- .SCF
- .QUAL
