Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (5): 92-102.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1322
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YAO Zi-ting1(), CAO Xue-ying2, XIAO Xue2, LI Rui-fang1, WEI Xiao-mei1, ZOU Cheng-wu2(), ZHU Gui-ning1()
Received:
2022-10-26
Online:
2023-05-26
Published:
2023-06-08
Contact:
ZOU Cheng-wu, ZHU Gui-ning
E-mail:youziting@163.com;zouchengwu@163.com;gnzhu@126.com
YAO Zi-ting, CAO Xue-ying, XIAO Xue, LI Rui-fang, WEI Xiao-mei, ZOU Cheng-wu, ZHU Gui-ning. Screening of Reference Genes for RT-qPCR in Neoscytalidium dimidiatum[J]. Biotechnology Bulletin, 2023, 39(5): 92-102.
序号 No. | 基因ID Gene ID | 引物 Primer | 引物序列 Primer sequence(5'-3') | 退火温度 Annealing temperature/℃ | 产物长度 PCR product length/bp | 基因名称 Gene name | 基因注释 Gene description |
---|---|---|---|---|---|---|---|
Gene1 | NEO_001463 | gene1-qF | GCGGTTTCCAGGTCTATCGTGAAG | 59 | 139 | CYT1 | Cytochrome c1 |
gene1-qR | GGCTCGGTGTCGTACTCGTTCT | ||||||
Gene2 | NEO_001659 | gene2-qF | CGTACTGGTGGAACTCGGAGGA | 58.5 | 151 | SDH2_1 | Succinate dehydrogenase subunit B |
gene2-qR | TTGAGAATGGTGTGGCAACGGTAA | ||||||
Gene3 | NEO_002342 | gene3-qF | GCTCGCCAATCTCCGTCGTT | 61.1 | 176 | TBCC | Tubulin-specific chaperone c |
gene3-qR | TCACCACCAGCACGCTATCCT | ||||||
Gene4 | NEO_002585 | gene4-qF | AGACACAGGCGAGACCAAGCA | 55.8 | 151 | SUI1 | Translation initiation factor SUI1 |
gene4-qR | GCAGGCAAACTTCTTCTTGATGACC | ||||||
Gene5 | NEO_003941 | gene5-qF | ATGCCGAGCCAGAAGTCTTTCC | 57.9 | 117 | RPL39 | Ribosomal L39 protein |
gene5-qR | GGCGTTGTACCTGATGGTGTTG | ||||||
Gene6 | NEO_004521 | gene6-qF | AAGTCTATGGCTTCTACGACGAGTG | 58.4 | 198 | PPH | Serine threonine-protein phosphatase pp2a catalytic subunit |
gene6-qR | GCGGCACCTCTTGTATCCTGTC | ||||||
Gene7 | NEO_004849 | gene7-qF | GCCACCAAGTTCGCTCCTATCC | 58.4 | 147 | ATP5B | ATP synthase F1 beta subunit |
gene7-qR | GGCACCACCGAAGAGACCAATC | ||||||
Gene8 | NEO_006464 | gene8-qF | CGGTGTCTTCTTCCTCGCTATCC | 58.3 | 183 | UBE2_16 | Ubiquitin-conjugating enzyme E2-16 kD |
gene8-qR | GAGCAGATGGAGAGCAGAACCTTC | ||||||
Gene9 | NEO_008113 | gene9-qF | TCACGAGTTCGGCTGGAAGTACC | 59.7 | 165 | RPL13 | Ribosomal L13 protein |
gene9-qR | TCGGCAAGCTGAGTCTTGACCTT | ||||||
Gene10 | NEO_008284 | gene10-qF | TCTGCTCAACGACCCGAACAC | 56.4 | 115 | UBE2_2 | Ubiquitin-conjugating enzyme e2_2 |
gene10-qR | CCAACTCTTCTCCACCGTCTCC | ||||||
Gene11 | NEO_008330 | gene11-qF | CTGTCAAGAAGTCCGCCAAGGT | 56.5 | 157 | RPS17 | Ribosomal protein S17e |
gene11-qR | CGCTTCATCAAGTGAGTGGTGTAG | ||||||
Gene12 | NEO_008873 | gene12-qF | TGGTGACATTGGCTGCCTGGT | 60.4 | 186 | SUCLA2 | Succinyl-CoA synthetase beta subunit |
gene12-qR | ATGTTGACGAAGATGGCGGTGAC | ||||||
Gene13 | NEO_010290 | gene13-qF | TCCTCTCATCTTCGCTGGTGTCA | 56.2 | 110 | ATP5A | ATP synthase F1 alpha subunit |
gene13-qR | GCTCGGACTCGTTGCTCTTCAG | ||||||
Gene14 | NEO_010575 | gene14-qF | CAAGGAGGATGCCGCCAACAA | 59 | 145 | TUB1_2 | Tubulin alpha-2 chain |
gene14-qR | ACCGCCGAAGGAGTGGAAGAT | ||||||
Gene15 | NEO_011231 | gene15-qF | CCAAGTCCAACCGTGAGAAGATGA | 56.7 | 102 | ACT1 | Actin |
gene15-qR | CGGAAGCGTACAGCGACAGAA | ||||||
Gene16 | NEO_011865 | gene16-qF | AGACCGACAACCGCCACTATGC | 60.1 | 184 | EFTU | Translation elongation factor Tu |
gene16-qR | ACAATCTTCTGGACACCGACCTGAC | ||||||
Gene17 | NEO_012815 | gene17-qF | CTGGTGAGTTCGAGGCTGGTATCT | 58.2 | 121 | EF1A | Translation elongation factor 1-alpha protein |
gene17-qR | CACTTGGTGGTGTCCATCTTGTTGA | ||||||
Gene18 | NEO_013111 | gene18-qF | CCGTACCGCTGCTACCAACATC | 59.8 | 195 | GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
gene18-qR | GCCTTGATAGTCGCCTTGATCTCAT |
Table 1 Primer sequences of candidate reference genes
序号 No. | 基因ID Gene ID | 引物 Primer | 引物序列 Primer sequence(5'-3') | 退火温度 Annealing temperature/℃ | 产物长度 PCR product length/bp | 基因名称 Gene name | 基因注释 Gene description |
---|---|---|---|---|---|---|---|
Gene1 | NEO_001463 | gene1-qF | GCGGTTTCCAGGTCTATCGTGAAG | 59 | 139 | CYT1 | Cytochrome c1 |
gene1-qR | GGCTCGGTGTCGTACTCGTTCT | ||||||
Gene2 | NEO_001659 | gene2-qF | CGTACTGGTGGAACTCGGAGGA | 58.5 | 151 | SDH2_1 | Succinate dehydrogenase subunit B |
gene2-qR | TTGAGAATGGTGTGGCAACGGTAA | ||||||
Gene3 | NEO_002342 | gene3-qF | GCTCGCCAATCTCCGTCGTT | 61.1 | 176 | TBCC | Tubulin-specific chaperone c |
gene3-qR | TCACCACCAGCACGCTATCCT | ||||||
Gene4 | NEO_002585 | gene4-qF | AGACACAGGCGAGACCAAGCA | 55.8 | 151 | SUI1 | Translation initiation factor SUI1 |
gene4-qR | GCAGGCAAACTTCTTCTTGATGACC | ||||||
Gene5 | NEO_003941 | gene5-qF | ATGCCGAGCCAGAAGTCTTTCC | 57.9 | 117 | RPL39 | Ribosomal L39 protein |
gene5-qR | GGCGTTGTACCTGATGGTGTTG | ||||||
Gene6 | NEO_004521 | gene6-qF | AAGTCTATGGCTTCTACGACGAGTG | 58.4 | 198 | PPH | Serine threonine-protein phosphatase pp2a catalytic subunit |
gene6-qR | GCGGCACCTCTTGTATCCTGTC | ||||||
Gene7 | NEO_004849 | gene7-qF | GCCACCAAGTTCGCTCCTATCC | 58.4 | 147 | ATP5B | ATP synthase F1 beta subunit |
gene7-qR | GGCACCACCGAAGAGACCAATC | ||||||
Gene8 | NEO_006464 | gene8-qF | CGGTGTCTTCTTCCTCGCTATCC | 58.3 | 183 | UBE2_16 | Ubiquitin-conjugating enzyme E2-16 kD |
gene8-qR | GAGCAGATGGAGAGCAGAACCTTC | ||||||
Gene9 | NEO_008113 | gene9-qF | TCACGAGTTCGGCTGGAAGTACC | 59.7 | 165 | RPL13 | Ribosomal L13 protein |
gene9-qR | TCGGCAAGCTGAGTCTTGACCTT | ||||||
Gene10 | NEO_008284 | gene10-qF | TCTGCTCAACGACCCGAACAC | 56.4 | 115 | UBE2_2 | Ubiquitin-conjugating enzyme e2_2 |
gene10-qR | CCAACTCTTCTCCACCGTCTCC | ||||||
Gene11 | NEO_008330 | gene11-qF | CTGTCAAGAAGTCCGCCAAGGT | 56.5 | 157 | RPS17 | Ribosomal protein S17e |
gene11-qR | CGCTTCATCAAGTGAGTGGTGTAG | ||||||
Gene12 | NEO_008873 | gene12-qF | TGGTGACATTGGCTGCCTGGT | 60.4 | 186 | SUCLA2 | Succinyl-CoA synthetase beta subunit |
gene12-qR | ATGTTGACGAAGATGGCGGTGAC | ||||||
Gene13 | NEO_010290 | gene13-qF | TCCTCTCATCTTCGCTGGTGTCA | 56.2 | 110 | ATP5A | ATP synthase F1 alpha subunit |
gene13-qR | GCTCGGACTCGTTGCTCTTCAG | ||||||
Gene14 | NEO_010575 | gene14-qF | CAAGGAGGATGCCGCCAACAA | 59 | 145 | TUB1_2 | Tubulin alpha-2 chain |
gene14-qR | ACCGCCGAAGGAGTGGAAGAT | ||||||
Gene15 | NEO_011231 | gene15-qF | CCAAGTCCAACCGTGAGAAGATGA | 56.7 | 102 | ACT1 | Actin |
gene15-qR | CGGAAGCGTACAGCGACAGAA | ||||||
Gene16 | NEO_011865 | gene16-qF | AGACCGACAACCGCCACTATGC | 60.1 | 184 | EFTU | Translation elongation factor Tu |
gene16-qR | ACAATCTTCTGGACACCGACCTGAC | ||||||
Gene17 | NEO_012815 | gene17-qF | CTGGTGAGTTCGAGGCTGGTATCT | 58.2 | 121 | EF1A | Translation elongation factor 1-alpha protein |
gene17-qR | CACTTGGTGGTGTCCATCTTGTTGA | ||||||
Gene18 | NEO_013111 | gene18-qF | CCGTACCGCTGCTACCAACATC | 59.8 | 195 | GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
gene18-qR | GCCTTGATAGTCGCCTTGATCTCAT |
Fig. 1 Electrophoresis analysis of total RNA M: 1 kb plus marker; 1-3: 3 biological repeats of control group; 4-6: 3 biological repeats of A group; 7-9: 3 biological repeats of B group; 10-12: 3 biological repeats of C group; 13-15: 3 biological repeats of D group
Fig. 2 Electrophoretic analysis of amplified products of 18 candidate reference genes M: 1 kb plus marker; nc: blank control; 1: CYT1; 2: SDH2_1; 3: TBCC; 4: SUI1;5: RPL39; 6: PPH; 7: ATP5B; 8: UBE2_16; 9: RPL13; 10: UBE2_2; 11: RPS17; 12: SUCLA2; 13: ATP5A; 14: TUB1_2; 15: ACT1; 16: EFTU; 17: EF1A; 18: GAPDH
序号 No. | 内参基因 Reference gene | 扩增效率 RT-qPCR efficiency/% | 相关系数 Correlation coefficient R2 | 序号 No. | 内参基因 Reference gene | 扩增效率 RT-qPCR efficiency/% | 相关系数 Correlation coefficient R2 | |
---|---|---|---|---|---|---|---|---|
Gene 1 | CYT1 | 106.4 | 0.998 5 | Gene 10 | UBE2_2 | 96.5 | 0.999 9 | |
Gene 2 | SDH2_1 | 106.2 | 0.986 5 | Gene 11 | RPS17 | 96.7 | 0.996 6 | |
Gene 3 | TBCC | 104.3 | 0.989 4 | Gene 12 | SUCLA2 | 103.6 | 0.996 6 | |
Gene 4 | SUI1 | 100.1 | 0.999 5 | Gene 13 | ATP5A | 101.9 | 0.996 9 | |
Gene 5 | RPL39 | 108.3 | 0.998 8 | Gene 14 | TUB1_2 | 93.6 | 0.999 6 | |
Gene 6 | PPH | 108.9 | 0.999 8 | Gene 15 | ACT1 | 91.6 | 0.999 9 | |
Gene 7 | ATP5B | 104.7 | 0.997 4 | Gene 16 | EFTU | 102.4 | 0.994 1 | |
Gene 8 | UBE2_16 | 104.2 | 0.999 9 | Gene 17 | EF1A | 94.8 | 0.996 8 | |
Gene 9 | RPL13 | 100.9 | 0.999 9 | Gene 18 | GAPDH | 90.9 | 0.995 0 |
Table 2 Amplification efficiencies of candidate reference genes
序号 No. | 内参基因 Reference gene | 扩增效率 RT-qPCR efficiency/% | 相关系数 Correlation coefficient R2 | 序号 No. | 内参基因 Reference gene | 扩增效率 RT-qPCR efficiency/% | 相关系数 Correlation coefficient R2 | |
---|---|---|---|---|---|---|---|---|
Gene 1 | CYT1 | 106.4 | 0.998 5 | Gene 10 | UBE2_2 | 96.5 | 0.999 9 | |
Gene 2 | SDH2_1 | 106.2 | 0.986 5 | Gene 11 | RPS17 | 96.7 | 0.996 6 | |
Gene 3 | TBCC | 104.3 | 0.989 4 | Gene 12 | SUCLA2 | 103.6 | 0.996 6 | |
Gene 4 | SUI1 | 100.1 | 0.999 5 | Gene 13 | ATP5A | 101.9 | 0.996 9 | |
Gene 5 | RPL39 | 108.3 | 0.998 8 | Gene 14 | TUB1_2 | 93.6 | 0.999 6 | |
Gene 6 | PPH | 108.9 | 0.999 8 | Gene 15 | ACT1 | 91.6 | 0.999 9 | |
Gene 7 | ATP5B | 104.7 | 0.997 4 | Gene 16 | EFTU | 102.4 | 0.994 1 | |
Gene 8 | UBE2_16 | 104.2 | 0.999 9 | Gene 17 | EF1A | 94.8 | 0.996 8 | |
Gene 9 | RPL13 | 100.9 | 0.999 9 | Gene 18 | GAPDH | 90.9 | 0.995 0 |
Fig. 3 Melting curve analysis of candidate reference genes Gene 1: CYT1; Gene 2: SDH2_1; Gene 3: TBCC; Gene 4: SUI1; Gene 5: RPL39; Gene 6: PPH; Gene 7: ATP5B; Gene 8: UBE2_16; Gene 9: RPL13; Gene 10: UBE2_2; Gene 11: RPS17; Gene 12: SUCLA2; Gene 13: ATP5A; Gene 14: TUB1_2; Gene 15: ACT1; Gene 16: EFTU; Gene 17: EF1A; Gene 18: GAPDH. The same below
Fig. 4 Expression levels of candidate reference genes in different culture conditions Boxes represent lower and upper quartiles of cycle thresholds range with medians indicated, and whisker caps represent maximum and minimum values
基因序号 No. | 基因名称 Gene name | 几何平均值 Geo mean | 算术平均值 AR mean | 最小值 Min | 最大值 Max | 相关系数 Correlation coefficient(r) | 标准偏差 Std dev | 变异系数 CV |
---|---|---|---|---|---|---|---|---|
Gene1 | CYT1 | 19.94 | 19.94 | 19.24 | 20.68 | 0.960 | 0.31 | 1.53 |
Gene2 | SDH2_1 | 19.69 | 19.69 | 19.06 | 20.16 | 0.754 | 0.30 | 1.54 |
Gene3 | TBCC | 22.90 | 22.92 | 21.50 | 24.66 | 0.915 | 0.83 | 3.63 |
Gene4 | SUI1 | 17.65 | 17.66 | 16.58 | 18.49 | 0.952 | 0.47 | 2.66 |
Gene5 | RPL39 | 19.71 | 19.72 | 18.77 | 20.67 | 0.851 | 0.63 | 3.21 |
Gene6 | PPH | 21.46 | 21.47 | 20.20 | 22.48 | 0.936 | 0.72 | 3.38 |
Gene7 | ATP5B | 18.21 | 18.22 | 17.97 | 18.74 | 0.521 | 0.20 | 1.11 |
Gene8 | UBE2_16 | 18.16 | 18.17 | 17.48 | 19.18 | 0.912 | 0.44 | 2.40 |
Gene9 | RPL13 | 19.97 | 19.98 | 18.81 | 20.69 | 0.761 | 0.56 | 2.79 |
Gene10 | UBE2_2 | 20.47 | 20.49 | 19.63 | 21.76 | 0.963 | 0.53 | 2.6 |
Gene11 | RPS17 | 19.82 | 19.83 | 18.73 | 20.78 | 0.803 | 0.61 | 3.09 |
Gene12 | SUCLA2 | 20.78 | 20.79 | 20.15 | 21.48 | 0.825 | 0.38 | 1.81 |
Gene13 | ATP5A | 19.49 | 19.50 | 18.88 | 20.03 | 0.87 | 0.33 | 1.67 |
Gene14 | TUB1_2 | 22.06 | 22.07 | 21.31 | 23.66 | 0.892 | 0.55 | 2.50 |
Gene15 | ACT1 | 21.30 | 21.33 | 19.76 | 22.72 | 0.835 | 1.13 | 5.29 |
Gene16 | EFTU | 21.48 | 21.49 | 20.66 | 22.36 | 0.577 | 0.65 | 3.00 |
Gene17 | EF1A | 16.04 | 16.05 | 15.35 | 16.48 | 0.893 | 0.31 | 1.96 |
Gene18 | GAPDH | 15.97 | 16.00 | 14.80 | 17.62 | 0.929 | 0.89 | 5.56 |
Table 3 Evaluating the expression stabilities of candidate reference genes by BestKeeper
基因序号 No. | 基因名称 Gene name | 几何平均值 Geo mean | 算术平均值 AR mean | 最小值 Min | 最大值 Max | 相关系数 Correlation coefficient(r) | 标准偏差 Std dev | 变异系数 CV |
---|---|---|---|---|---|---|---|---|
Gene1 | CYT1 | 19.94 | 19.94 | 19.24 | 20.68 | 0.960 | 0.31 | 1.53 |
Gene2 | SDH2_1 | 19.69 | 19.69 | 19.06 | 20.16 | 0.754 | 0.30 | 1.54 |
Gene3 | TBCC | 22.90 | 22.92 | 21.50 | 24.66 | 0.915 | 0.83 | 3.63 |
Gene4 | SUI1 | 17.65 | 17.66 | 16.58 | 18.49 | 0.952 | 0.47 | 2.66 |
Gene5 | RPL39 | 19.71 | 19.72 | 18.77 | 20.67 | 0.851 | 0.63 | 3.21 |
Gene6 | PPH | 21.46 | 21.47 | 20.20 | 22.48 | 0.936 | 0.72 | 3.38 |
Gene7 | ATP5B | 18.21 | 18.22 | 17.97 | 18.74 | 0.521 | 0.20 | 1.11 |
Gene8 | UBE2_16 | 18.16 | 18.17 | 17.48 | 19.18 | 0.912 | 0.44 | 2.40 |
Gene9 | RPL13 | 19.97 | 19.98 | 18.81 | 20.69 | 0.761 | 0.56 | 2.79 |
Gene10 | UBE2_2 | 20.47 | 20.49 | 19.63 | 21.76 | 0.963 | 0.53 | 2.6 |
Gene11 | RPS17 | 19.82 | 19.83 | 18.73 | 20.78 | 0.803 | 0.61 | 3.09 |
Gene12 | SUCLA2 | 20.78 | 20.79 | 20.15 | 21.48 | 0.825 | 0.38 | 1.81 |
Gene13 | ATP5A | 19.49 | 19.50 | 18.88 | 20.03 | 0.87 | 0.33 | 1.67 |
Gene14 | TUB1_2 | 22.06 | 22.07 | 21.31 | 23.66 | 0.892 | 0.55 | 2.50 |
Gene15 | ACT1 | 21.30 | 21.33 | 19.76 | 22.72 | 0.835 | 1.13 | 5.29 |
Gene16 | EFTU | 21.48 | 21.49 | 20.66 | 22.36 | 0.577 | 0.65 | 3.00 |
Gene17 | EF1A | 16.04 | 16.05 | 15.35 | 16.48 | 0.893 | 0.31 | 1.96 |
Gene18 | GAPDH | 15.97 | 16.00 | 14.80 | 17.62 | 0.929 | 0.89 | 5.56 |
序号 No. | 基因 Genes | geNorm | NormFinder | BestKeeper | RefFinder | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
M值 M value | 排序 Rank | 稳定值 Stability value | 排序 Rank | 变异度 CV/% | 标准差 SD | 相关系数 Correlation coefficientr | 排序 Rank | 稳定性 Stability value | 综合排序 Comprehensive rank | ||||||
Gene1 | CYT1 | 0.171 | 2 | 0.186 | 2 | 1.53 | 0.31 | 0.960 | 1 | 2.06 | 1 | ||||
Gene2 | SDH2_1 | 0.245 | 3 | 0.383 | 9 | 1.54 | 0.30 | 0.754 | 13 | 5.05 | 7 | ||||
Gene3 | TBCC | 0.509 | 15 | 0.563 | 15 | 3.63 | 0.83 | 0.915 | 7 | 15.49 | 15 | ||||
Gene4 | SUI1 | 0.306 | 5 | 0.185 | 1 | 2.66 | 0.47 | 0.952 | 3 | 3.13 | 2 | ||||
Gene5 | RPL39 | 0.393 | 9 | 0.391 | 10 | 3.21 | 0.63 | 0.851 | 10 | 10.68 | 11 | ||||
Gene6 | PPH | 0.457 | 13 | 0.392 | 11 | 3.38 | 0.72 | 0.936 | 5 | 12.63 | 14 | ||||
Gene7 | ATP5B | 0.436 | 12 | 0.499 | 14 | 1.11 | 0.2 | 0.521 | 16 | 7.10 | 9 | ||||
Gene8 | UBE2_16 | 0.333 | 6 | 0.232 | 4 | 2.40 | 0.44 | 0.912 | 4 | 5.29 | 8 | ||||
Gene9 | RPL13 | 0.420 | 11 | 0.462 | 13 | 2.79 | 0.56 | 0.761 | 14 | 12.22 | 13 | ||||
Gene10 | UBE2_2 | 0.353 | 7 | 0.225 | 3 | 2.60 | 0.53 | 0.963 | 2 | 5.05 | 6 | ||||
Gene11 | RPS17 | 0.408 | 10 | 0.449 | 12 | 3.09 | 0.61 | 0.803 | 15 | 11.74 | 12 | ||||
Gene12 | SUCLA2 | 0.159 | 1 | 0.312 | 7 | 1.81 | 0.38 | 0.825 | 12 | 4.14 | 4 | ||||
Gene13 | ATP5A | 0.159 | 1 | 0.286 | 6 | 1.67 | 0.33 | 0.870 | 8 | 3.50 | 3 | ||||
Gene14 | TUB1_2 | 0.372 | 8 | 0.341 | 8 | 2.50 | 0.55 | 0.892 | 9 | 8.71 | 10 | ||||
Gene15 | ACT1 | 0.576 | 17 | 0.839 | 18 | 5.29 | 1.13 | 0.835 | 18 | 18.00 | 18 | ||||
Gene16 | EFTU | 0.485 | 14 | 0.622 | 17 | 3.00 | 0.65 | 0.577 | 17 | 15.70 | 16 | ||||
Gene17 | EF1A | 0.272 | 4 | 0.276 | 5 | 1.96 | 0.31 | 0.893 | 6 | 4.95 | 5 | ||||
Gene18 | GAPDH | 0.533 | 16 | 0.588 | 16 | 5.56 | 0.89 | 0.929 | 11 | 16.49 | 17 |
Table 4 Analyzing the expression stabilities of genes by GeNorm, NormFinder, BestKeeper and RefFinder
序号 No. | 基因 Genes | geNorm | NormFinder | BestKeeper | RefFinder | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
M值 M value | 排序 Rank | 稳定值 Stability value | 排序 Rank | 变异度 CV/% | 标准差 SD | 相关系数 Correlation coefficientr | 排序 Rank | 稳定性 Stability value | 综合排序 Comprehensive rank | ||||||
Gene1 | CYT1 | 0.171 | 2 | 0.186 | 2 | 1.53 | 0.31 | 0.960 | 1 | 2.06 | 1 | ||||
Gene2 | SDH2_1 | 0.245 | 3 | 0.383 | 9 | 1.54 | 0.30 | 0.754 | 13 | 5.05 | 7 | ||||
Gene3 | TBCC | 0.509 | 15 | 0.563 | 15 | 3.63 | 0.83 | 0.915 | 7 | 15.49 | 15 | ||||
Gene4 | SUI1 | 0.306 | 5 | 0.185 | 1 | 2.66 | 0.47 | 0.952 | 3 | 3.13 | 2 | ||||
Gene5 | RPL39 | 0.393 | 9 | 0.391 | 10 | 3.21 | 0.63 | 0.851 | 10 | 10.68 | 11 | ||||
Gene6 | PPH | 0.457 | 13 | 0.392 | 11 | 3.38 | 0.72 | 0.936 | 5 | 12.63 | 14 | ||||
Gene7 | ATP5B | 0.436 | 12 | 0.499 | 14 | 1.11 | 0.2 | 0.521 | 16 | 7.10 | 9 | ||||
Gene8 | UBE2_16 | 0.333 | 6 | 0.232 | 4 | 2.40 | 0.44 | 0.912 | 4 | 5.29 | 8 | ||||
Gene9 | RPL13 | 0.420 | 11 | 0.462 | 13 | 2.79 | 0.56 | 0.761 | 14 | 12.22 | 13 | ||||
Gene10 | UBE2_2 | 0.353 | 7 | 0.225 | 3 | 2.60 | 0.53 | 0.963 | 2 | 5.05 | 6 | ||||
Gene11 | RPS17 | 0.408 | 10 | 0.449 | 12 | 3.09 | 0.61 | 0.803 | 15 | 11.74 | 12 | ||||
Gene12 | SUCLA2 | 0.159 | 1 | 0.312 | 7 | 1.81 | 0.38 | 0.825 | 12 | 4.14 | 4 | ||||
Gene13 | ATP5A | 0.159 | 1 | 0.286 | 6 | 1.67 | 0.33 | 0.870 | 8 | 3.50 | 3 | ||||
Gene14 | TUB1_2 | 0.372 | 8 | 0.341 | 8 | 2.50 | 0.55 | 0.892 | 9 | 8.71 | 10 | ||||
Gene15 | ACT1 | 0.576 | 17 | 0.839 | 18 | 5.29 | 1.13 | 0.835 | 18 | 18.00 | 18 | ||||
Gene16 | EFTU | 0.485 | 14 | 0.622 | 17 | 3.00 | 0.65 | 0.577 | 17 | 15.70 | 16 | ||||
Gene17 | EF1A | 0.272 | 4 | 0.276 | 5 | 1.96 | 0.31 | 0.893 | 6 | 4.95 | 5 | ||||
Gene18 | GAPDH | 0.533 | 16 | 0.588 | 16 | 5.56 | 0.89 | 0.929 | 11 | 16.49 | 17 |
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