5
TAKARA BIO INC.
URL:http://www.takara-bio.com
CycleavePCR
TM
O157 (VT1/VT2) Detection Kit Ver.2.0
v.0607
Cat.#CY203
[Preparation of heat-extracted bacterial sample]
1) Transfer 10
µ
l of proliferated bacterial culture into a 1.5 ml tube.
2) Add 90
µ
l of sterilized water and mix.
3) Heat for 5 min. at 95
°
C.
4) Centrifuge at 12,000 rpm, 4
°
C for 10 min, and collect the supernatant. Use the obtained
supernatant as heat-extracted sample for VT1/VT2 detection.
* If the PCR reaction is inhibited with heat-extracted sample prepared through the above
method, dilute it with sterilized water by 10-fold and 100-fold and apply them for PCR
reaction.
* Proliferated culture solution should be prepared by following the standard protocol
appropriate for each food sample. Heat extracted sample can be stored at -20
°
C
.
(For more information on handling Smart Cycler
®
II System, see the instructions supplied with it.)
(1) Start the Smart Cycler
®
II System.
(2) Set the protocol.
Click the icon “Define Protocols” and then “New Protocol” button to create the protocol
by following the steps shown below. (Since the created protocol is saved, no entry is
required in subsequent reactions).
VI-1. Sample preparation (Perform in Area 2)
VI-2.
Setting of Smart Cycler
®
II System (Perform in Area 3)
↓
Enter sample name. [Sample ID]
Set analytic parameters. [Analysis Setting]
↓
Amplification curve is viewed on the screen in real time.
↓
The reaction is terminated.
↓
5. Judgement (see page 12-16).
Define Protocols
Crick New Protocol
Create this protocol