15
TAKARA BIO INC.
URL:http://www.takara-bio.com
CycleavePCR
TM
O157 (VT1/VT2) Detection Kit Ver.2.0
v.0607
Cat.#CY203
*1. The VT1 or VT2 gene is determined to be positive whether the result of Internal
Control is POS or NEG. Confirm that there is no contamination in the reaction referring to
the result of negative control reaction.
*2. Confirm that a positive control reaction is proved to be POS. That is, there is no
contamination in the reaction.
*3. The PCR reaction or cycling probe detection process was not be correctly performed for
some reason. Retry the reaction process. Substances that inhibit reaction can be included
in the sample. Repreparation of sample may be required.
*4. It is suspected that there may be problem in Primers for VT1 amplification or in Probe
for VT1 detection, or that VT1 Positive Control may be degraded.
*5. It is suspected that there may be problem in Primers for VT2 amplification or in Probe
for VT2 detection, or that VT2 Positive Control may be degraded.
*6. Contamination is suspected. Retry after decontamination of the place where the reaction
mixture is prepared and the equipment used.
VII. Trouble shooting
(1) A numeric value appears in the FAM,TET, or TxR Std/Res column instead of POS or NEG.
→
When Sample Type has been set as STD in Results Table, a numeric value
appears.
All the data in the Sample Type must be set as UNKN.
(2) POS appears in the FAM Std/Res ,TET Std/Res, or ROX Std/Res column accidentally
due to, for example, the signal noises other than those from the amplified product.
→
Make judgement based on the shape of the amplified curve. If the judgement is
difficult, retry the reaction process.
1
1
1
10
0
0
00
0
0
0
1
1
1
10
0
0
00
0
0
0
Amplified curve derived from
an amplified product
(Sigmoid curve is drawn.)
Increased signal due to any
other reason than an amplified
product