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Microbial Identification reagents

Click to Read Overview (below products)
Product Id (Sort)Description (Sort)PDFSize/Vol.$ (U.S.)QtyAdd/Update
SEQ16S-KITBacteria Ribosome 16S Sequencing Kit [includes 4 PCR Master Mix (SEQ16S-MM, 1 Primers (SEQ16S-P)] PDF  $210.00
SEQ16S-MMBacteria Ribosome 16S Sequencing PCR Master Mix PDF 0.350ml$50.00
SEQ16S-PBacteria Ribosome 16S Sequencing Primer (10uM) PDF 0.500ml$50.00
SEQITS-KITFungal ITS Sequencing Kit [includes 4 PCR Master Mix (SEQITS-MM) , 1 Primers (SEQITS-P), 1 Bead (SEQITS-B)] PDF  $210.00
SEQITS-MM Fungal ITS Sequencing Master Mix PDF 0.350ml$50.00
SEQITS-PFungal ITS Sequencing Primer (10uM) PDF 0.500ml$50.00
SEQITS-B15Fungal ITS Sequencing Beads (0.5mm dia) PDF 30gm (15: 2ml vials)$75.00
SEQ16S-RBacterial Ribosome 16S Sequencing (Reverse) Primer (10uM) PDF 0.500ml$50.00
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Overview   top ^

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.



1: Applied Biosystems
2: USB Corporation
Attostar LLC