Direct Colony Sequencing
In order to reduce your routine lab workload, ACGT, Inc. offers Direct Colony Sequencing Service. Instead of purified plasmid DNA, just send us bacterial colonies re-suspended in a small volume of buffer and a sequencing primer (separately or in the same tube). Our skilled professionals will extract and amplify the plasmid DNA directly from the colony, and generate sequencing data with the same exceptional level of quality as our standard sequencing service.
Sample Preparation and Submission
- Pick a bacterial colony from an overnight plate, or pipette 1 ul of overnight bacterial culture, phage lysate, or glycerol stock.
- Deposit the bacterial culture aliquot into a labeled Eppendorf tube containing 50 ul of a buffered saline. We recommend PBS pH 7.2 to 7.5, or TES (TrisHCl 10mM, EDTA 1mM, NaCl 150mM) pH 7.2 to 7.5. DO NOT use distilled water.
- Vortex briefly to re-suspend.
- Ship to ACGT, Inc. at ambient temperature using our Drop Box service or any overnight shipping service.
Direct Colony Method Works on Plasmids, Cosmids, BACs, PACs and Phages
Any size plasmid can be sequenced by this method, and it works equally well with high copy or low copy plasmid templates. Typically, large template sequencing (cosmids, BACs and PACs) is difficult and requires lengthy and laborious template preparation – with no guarantee that good quality sequencing results will be obtained. With our Direct Colony Sequencing Service, high quality sequences are routinely generated from the templates even up to the size of BACs and PACs. We can also generate sequences from phage lysates.
Direct Colony Sequencing Service is ideal for screening cDNA libraries or SDM products. As an extra benefit to you, we will keep the original bacterial sample tubes that you’ve submitted for up to 2 weeks, which gives you an option to request plasmid DNA preparation service (at any scale of your choice) or site-directed mutagenesis (SDM) service without the extra expense or time delay of shipping the sample to ACGT, Inc. Just follow the link provided with your sequencing results to indicate which of the samples you have submitted requires DNA preparation or SDM services.