Direct Colony Sequencing

Sanger sequencing of plasmid DNA extracted and amplified from bacterial colonies or culture in buffer.

Eliminate the need for plasmid preparation with our direct colony sequencing service. Obtain accurate and reliable genetic data directly from bacterial colonies.

Service Description

Extraction and amplification of plasmid DNA from colonies or culture in buffer
Sanger sequencing reactions performed using BigDye terminator
Cleanup with magnetic beads for superior quality traces
Analysis of the extension products using our ABI 3730 Genetic Analyzer
Data review and troubleshooting for problematic samples

Sample Preparation and Submission

Pick

Pick a bacterial colony from an overnight plate, or pipette 1 ul of overnight bacterial culture, phage lysate, or glycerol stock.

Deposit

Add to a labeled microfuge tube containing 50 ul of a buffered saline.
PBS pH 7.2 to 7.5
TES (TrisHCl 10mM, EDTA 1mM, NaCl 150mM) pH 7.2 to 7.5
Do not use distilled water.

Vortex

Vortex briefly to re-suspend.

Ship

Ship to ACGT at ambient temperature using our dropbox service or any overnight shipping service.

Turnaround Time

Initial results will be available 2-3 business days after receiving your samples. Reactions that require additional troubleshooting may result in longer turnaround times.

Acceptable DNA Types

Plasmids
Cosmids
BACs
PACs
Phages

Any size plasmid can be sequenced by Direct Colony Sequencing, and it works just as well with high copy or low copy plasmid templates. Typically, large template sequencing (cosmids, BACs and PACs) can be difficult. Moreover, it requires a lengthy template preparation without an assurance of obtaining high quality sequencing results. With our Direct Colony Sequencing Service, high quality sequences are routinely generated from templates, even up to the size of BACs and PACs. We can also generate sequences from phage lysates.

Highlights

Generates 750 to 1,000 bases of data
Extensive troubleshooting for unsuccessful samples
Repeat reactions provided at no additional cost
Free primer banking and template storage by request
Free use of ACGT universal primers

Does your order contain 48 reactions or more?

For high volume samples, please submit your order through our High Throughput service. All aspects, including sample preparation, alternative protocol selection, and data quality remain unchanged, with the added benefit of discounted rates for larger volumes. Samples must be submitted in 96-well plates or PCR strip tubes to qualify.

Go to HTP: Sanger Sequencing

Does your order consist of 48 reactions or more?

For high volume samples, please submit your order through our High-Throughput service. All aspects, including sample preparation, alternative protocol selection, and data quality remain unchanged, with the added benefit of discounted rates for larger volumes. Samples must be submitted in 96-well plates or PCR strip tubes to qualify.