INMV Typing Method
List of primers
| TR no. |
Position of primer on M. avium subsp. paratuberculosis genome
|
Predicted size of PCR product (bp) |
Primer
|
Tma (°C) | Buffer usedb
|
| Start |
Stop |
Forward |
Reverse |
1 μl DMSO |
5 μl betaine |
| 292 |
3253590 |
3253889 |
300 |
CTTGAGCAGCTCGTAAAGCGT |
GCTGTATGAGGAAGTCTATTCATGG |
58 |
+ |
− |
| X3 |
4441875 |
4442070 |
196 |
AACGAGAGGAAGAACTAAGCCG |
TTACGGAGCAGGAAGGCCAGCGGG |
58 |
− |
− |
| 25 |
3665598 |
3665947 |
350 |
GTCAAGGGATCGGCGAGG |
TGGACTTGAGCACGGTCAT |
58 |
+ |
− |
| 47 |
4128604 |
4128821 |
217 |
CGTTGCGATTTCTGCGTAGC |
GGTGATGGTCGTGGTCATCC |
64 |
+ |
− |
| 3 |
131320 |
131527 |
208 |
CATATCTGGCATGGCTCCAG |
ATCGTGTTGACCCCAAAGAAAT |
60 |
+ |
− |
| 7 |
3711417 |
3711619 |
203 |
GACAACGAAACCTACCTCGTC |
GTGAGCTGGCGGCCTAAC |
60 |
+ |
− |
| 10 |
4279553 |
4279855 |
303 |
GACGAGCAGCTGTCCGAG |
GAGAGCGTGGCCATCGAG |
60 |
− |
+ |
| 32 |
1125707 |
1126004 |
298 |
CCACAGGGTTTTTGGTGAAG |
GGAAATCCAACAGCAAGGAC |
55 |
+ |
+ |
aTm, melting temperature.
bDMSO, dimethyl sulfoxide. +, used; −, not used. (PCR reaction in 25 µL)
Schematic representation with genome of K10
