8
TAKARA BIO INC.
CycleavePCR
TM
O157 (VT1/VT2) Detection Kit Ver.2.0
v.0607
Cat.#CY203
URL:http://www.takara-bio.com
Per reaction final conc.
5 X Reaction Mixture 5
µ
l 1 X
VT Primer Mix (5
µ
l each) 2
µ
l 0.4
µ
M each
VT Chimera Probe Mix (5 X conc.) 5
µ
l 1 X
TaKaRa Ex Taq R-PCR (5 units/
µ
l) 0.25
µ
l 1.25 units
Tli RNase H II (200 units/
µ
l) 0.5
µ
l 100 units
Sample DNA or Positive Control 1~10
µ
l
Sterilize distilled water up to 25
µ
l
Sample (template) solution is added in the volume range of 1-10
µ
l. Add sterilized distilled
water to have the final volume of 25
µ
l. In case of adding positive control, add 1
µ
l as
template.
Add the components of the above reaction mixture without template into a fresh tube, by
pouring on the tube wall. Please refer to the following figure.
Close gently the lid of the tubes not so tightly and move to the Area 3.
(2) Addition of sample template (Perfom in Area 3):
Prepare one tube of negative control by adding sterilized distilled water instead of sample.
For the rest tubes, add the prepared samples into the reaction mixture prepared at (1),
and close the lid of the tube tightly.
Place the reaction mixture (15-24
µ
l)
into this reserver part.
Centrifuge to load down the reaction
mixture into this part of the tube.
(3) Load the reaction tubes on Smart Cycler
®
II System and click the "Start Run" button to start
the reaction process.