DIRECT COLONY SEQUENCING
In effort to reduce your lab workload, ACGT offers our Direct Colony Sequencing Service (DCSS). Instead of purified plasmid DNA, simply send us bacterial colonies re-suspended in a small volume of buffer with a sequencing primer in a separate tube. Our skilled professionals will extract and amplify the plasmid DNA directly from the colony and generate sequencing data with the same high quality as our standard sequencing service.
Sample Preparation and Submission
1. Pick a bacterial colony from an overnight plate, or pipette 1 ul of overnight bacterial culture, phage lysate, or glycerol stock.
2. 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.
3. Vortex briefly to re-suspend.
4. Ship to ACGT at ambient temperature using our dropbox service or any overnight shipping service.
DCSS Works on Plasmids, Cosmids, BACs, PACs and 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.
Direct Colony Sequencing Service is ideal for screening cDNA libraries or SDM products. As an added benefit, ACGT will maintain the original bacterial sample tubes that you submitted for up to 2 weeks, which provides you with the option to request our 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 us.