| Name |
Sequence (5' to 3')* |
|
| Sense minigene |
|
| Forward primer |
CAG AGA TGC ATA ATA CGA CTC ACT ATA GGG AGA AGG GTG TGC GGC GTG CAC
TGG ACT TT |
| Reverse primer |
AGG GTG TGC GGC GTG CAC TGG ACT TT |
| Antisense minigene |
|
| Forward primer |
CAG AGA TGC ATA ATA CGA CTC ACT ATA GGG AGA GGT CAT GGA AGG GGC AGT
TGT CCA AGT T |
| Reverse primer |
GGT CAT GGA AGG GGC AGT TGT CCA AGT T |
| QRT-PCR primers |
|
| MA097 |
CGG CGA GTA CAA CAA AGC CA |
| MA098 |
CAC AGC GTA GAT CTG GAA AG |
| QRT-PCR probe |
|
| MA576 |
(FAM) -CTC CAC GTC CAA GAA GTA GTT CA- (TAMRA) |
|
| *The sequences of oligonucleotide primers used for polymerase chain reaction
(PCR) amplification to construct cystatin C minigenes are shown with the T7
RNA promoter sequence underlined. These minigenes were used for riboprobe
synthesis with T7 RNA polymerase. To quantitate cystatin C messenger RNA,
complementary DNA was generated from total RNA by reverse transcriptase and
random hexamer primers, after which cystatin C sequences were specifically
amplified by quantitative real-time PCR (QRT-PCR) with the use of primers
MA097 and MA098. The cystatin C-derived product was measured in real-time by
the TaqMan probe MA576, with a 5' fluorescent reporter (FAM) and a
3' quencher (TAMRA) |
