SAMPLE PREPARATION

 

DNA is isolated from samples by the investigator. Do not send actual clinical samples unless special arrangements are made in advance. For technical 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.

 

The 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 from the hard-to-lyse oral species. Clearly, if there is not adequate lysis, then species not lysed will not be detected using HOMINGS.

 

Sample Submission

 

Each DNA sample must be adjusted to a concentration of 20 ng/µl and at least 10 µl of each sample must be provided. Measurements are typically made using a NanoDrop (Thermo Scientific). Samples must also have a 260/280 ratio of 1.8 or greater. DNA samples in solution should be frozen at -80°C. Lyophilization of samples is recommended for international clients.

 

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

 

All samples must be clearly labeled without patient identifiers.

Samples will be screened initially for suitability for HOMINGS processing (see pricing).

 

Once approved, ship samples to:
Alexis Kokaras (HOMINGS)
The Forsyth Institute
245 First Street
Cambridge, MA 02142