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THE PROBLEM:
Some bacteria, yeasts and fungi take too long, or are too difficult, to identify in the clinical microbiology laboratory. After 3 days, the struggle needs to be overcome with effiency.

THE ANSWER:
Sequence a region of the 16S ribosome chromosome site for bacteria or the ITS chromosome site for yeast and fungi. Attostar provides reagents and procedures to get this done with a minimum of cost and effort:

The reagents:
16S Master Mix,
16S Sequencing primer,
ITS Master Mix,
ITS Sequencing primer.

Take advantage of two very nice new developments:

• First, NCBI Blast results now come with a great visual tree for results:
• Second, DNA sequencing capabilities have tremendous excess capacity.

The procedure:

• Make a suspension of the unknown bacteria, yeast, or mold in PrepMan® Ultra Sample Preparation Reagent¹.
• Release the bacterial DNA by heating at 100ºC for 10 minutes.
• Release yeast and fungal DNA by vortexing with 0.5mm glass beads (provided) followed by heating at 100ºC for 10 minutes.
• Dilute the DNA.
• Use the DNA in an Attostar PCR master mix. Add AmpliTaq Gold® LD¹. Do PCR.
• Treat the PCR product with ExoSAP-IT®² at 37ºC 10 minutes and 80ºC for 10 minutes.
• Send the resulting mixture (and Attostar primer) to a DNA sequencing facility of your choice. (There are many available that will do a sequence for $8. The results are usually sent back the same day.)
• Copy and paste the results into the NCBI Blast site, and
• A few seconds later your results are available.