About this article
Article Category: original-paper
Published Online: Sep 04, 2018
Page range: 347 - 353
Received: Mar 19, 2018
Accepted: Jun 14, 2018
DOI: https://doi.org/10.21307/pjm-2018-042
Keywords
© 2018 Takema Hasegawa et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Fig. 1.
Fig. 2.
Fig. 3.
Primers used in the RT-qPCR.
| Target | Forward Primer | Reverse Primer |
|---|---|---|
| 1000-A | 5’-CAACCGGTGTGATCAGGACA-3’ | 5’-AGGACAGTCCGCATAAGCAC-3’ |
| 1000-B | 5’-TACCAGCGCTTCTGTACGAC-3’ | 5’-GAGCTGTATCCGTGCCGTAA-3’ |
| 500-A | 5’-TCGCAGGCCTAATACGTGTC-3’ | 5’-CGTGAATCTCGGAGCGGTAA-3’ |
| 500-B 3’end | 5’-GGGTAGCGATTTAACGACTCG-3’ | 5’-CAGAGCCTGCCTTATCGTGA-3’ |
| 500-B middle | 5’-CCGAACGCTACGTGACGATA-3’ | 5’-ATCTACATGTTCCGTGCGCA-3’ |
| 500-B 5’end | 5’-AGACTAAATCTCGGCGTCGG-3’ | 5’-TAGATAGGGTCCGCATGACG-3’ |
| 500-C 3’end | 5’-GCACGACCGAATTATGCACC-3’ | 5’-AACCACTGACGTGAGCGATT-3’ |
| 500-C middle | 5’-TAGACGCGCCTTACTCCTCT-3’ | 5’-TAGTGGAGCTCGCGGATTTG-3’ |
| 500-C 5’end | 5’-GGACTAAACGCACTGAATACCG-3’ | 5’-ATCGCCCGTACTATCCGGTA-3’ |
Inhibition of RT-qPCR by RNA extract solutions from S. cerevisiae and E. coli were evaluated using standard RNAs (%).
| Species | 1000-A | 1000-B | 500-A |
|---|---|---|---|
| –1.1 ± 4.8 | 50.4 ± 4.0 | 0.3 ± 3.1 | |
| 7.0 ± 3.2 | 44.6 ± 3.0 | 27.7 ± 2.8 |
Survival of 3’ end and 5’ end and the ratio of 3’ end to 5’ end regions of standard RNAs after degradation with S. cerevisiae RNA crude extract.
| Standard RNA | 500-B | 500-C | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Degradation time (min) | 0 | 10 | 20 | 60 | 180 | 0 | 10 | 20 | 60 | 180 |
| 3’ end (survival %) | 100 | 73.5 | 45.2 | 32.1 | 6.3 | 100 | 76 | 59.2 | 26.7 | 3.2 |
| 5’ end (survival %) | 100 | 85.9 | 71.2 | 67.9 | 6.4 | 100 | 88.9 | 92.9 | 67.2 | 8.6 |
| 3’ end /5’ end (ratio) | 1 | 0.86 | 0.63 | 0.47 | 0.99 | 1 | 0.86 | 0.64 | 0.4 | 0.37 |