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REVISED 121213
(e)
Pipettor
. – 100-1,000 µL, adjustable volume.
1
(f)
8-channel micropipettor
. – 50 µL, fixed or adjustable volume.
2
(g)
Pipette tips
. – 50 µL, with filter.
3
(h)
Pipette tips
. – 1,000 µL.
4
(i)
Tubes.
– Glass or plastic, 12 × 75 mm or similar, sterile, with caps.
5
(j)
Inoculating loops or needles
. – Sterile.
6
D.
Preparation of Test Samples
7
Pick an isolated colony from non-selective or selective/differential agar medium (one of the media listed
8
in section B) with an inoculating loop or needle and resuspend (vortex or otherwise thoroughly mix) in
9
0.5 mL PBS in a sterile, capped tube.
10
E.
Test Procedure
11
General Preparation:
12
(a)
This assay should be performed in a controlled laboratory environment.
13
(b)
Do not use culture media or ANSR reagents beyond their expiration dates. Do not interchange
14
reagents between ANSR kit lots.
15
(c)
Remove ANSR reaction tubes from the foil pouch just before use. Avoid prolonged exposure to
16
light. Tap reaction tubes on bench top to make sure that lyophilized reagents are at the bottom of the
17
tube prior to adding the lysed sample.
18
(d)
Complete all assay steps in sequence, avoiding delays between steps.
19
(e)
Exercise care in pipetting steps to avoid cross-contamination of samples.
20
(f)
Do not remove caps from reaction tubes at any point after the assay is started
; this will
21
prevent accidental contamination of the environment with amplification products.
22
(g)
Prior to starting the assay:
23
(i)
Preheat the lysis heater block to 80 ± 2
o
C.
24
7