SAMPLE PREPARATION

DNA is isolated from clinical samples by the client and the DNA samples are then submitted. Thus, do not send actual clinical samples unless special arrangements are made in advance. For technicial assistance, contact Alexis Kokaras at akokaras@forsyth.org or (617) 892-8567.

Recommended DNA isolation protocol. The following procedure is suitable for isolation of DNA from hard-to-lyse species, such as S. mutans, Staphylococcus spp., Actinomyces spp., and Scardovia spp. This procedure works well for samples from all oral sites including saliva, supragingival or subgingival plaque, buccal epithelial cells, scrapings from tongue surfaces, palatal and throat swabs, and denture surfaces.

Modified Protocol uses Ready-Lyse™ Lysozyme Solution (Epicentre, Cat. No. R1802M) for overnight incubation and MasterPure DNA Purification Kit (Epicentre, Cat. No. MCD85201)

Please call for advice if you intend to use other protocols. Note that some of these protocols may not be sufficient for DNA isolation of the hard-to-lyse oral species. Clearly, if there is not adequate lysis, then those "unlysed" species will not be detected using HOMIM.

Sample Submission

At least 200 ng at a concentration of a minimum of 20 ng/µl of each genomic DNA sample in a 0.5 ml or 1.5 ml Eppendorf tube must be submitted. However, samples at higher concentrations are preferred. Samples must have a 260/280 ratio of 1.6-2.0. DNA samples in solution should be frozen at -80°C. Lyophilization of samples is recommended for international clients.

All frozen samples must be shipped overnight on dry ice (note there are no weekend or holiday deliveries to the Forsyth Institute). Lyophilized samples can be shipped at room temperature.

All samples must be clearly labeled without patient identifiers.
Samples will be initially screened for suitability for HOMIM processing (see pricing).

Once approved by MIM, ship samples to:

Alexis Kokaras (HOMIM)
The Forsyth Institute
245 First Street
Cambridge, MA 02142