Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (9): 141-147.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0611
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YUAN Lan1,2(), HUANG Ya-nan2, ZHANG Bei-ni1,2, XIONG Yu-meng1, WANG Hong-yang1()
Received:
2024-07-01
Online:
2024-09-26
Published:
2024-10-12
Contact:
WANG Hong-yang
E-mail:2508701413@qq.com;hongyang8318@ynnu.edu.cn
YUAN Lan, HUANG Ya-nan, ZHANG Bei-ni, XIONG Yu-meng, WANG Hong-yang. High-throughput Sample Preparation Method for the Identification of Potato Ploidy Using Flow Cytometry[J]. Biotechnology Bulletin, 2024, 40(9): 141-147.
样品Sample | 浓度Concentration | 分子量 Molecular weight | 添加量 Additive amount |
---|---|---|---|
MgCl2·6H2O | 0.045 mol/L | 203.3 | 4.574 g |
Na3C6H5O7·2H2O | 0.030 mol/L | 294.1 | 4.411 g |
C7H15NO4S | 0.020 mol/L | 209.27 | 2.093 g |
Triton X-100 | 0.1%(体积比) | 0.5 mL |
Table 1 Preparation of 500 mL lysis buffer
样品Sample | 浓度Concentration | 分子量 Molecular weight | 添加量 Additive amount |
---|---|---|---|
MgCl2·6H2O | 0.045 mol/L | 203.3 | 4.574 g |
Na3C6H5O7·2H2O | 0.030 mol/L | 294.1 | 4.411 g |
C7H15NO4S | 0.020 mol/L | 209.27 | 2.093 g |
Triton X-100 | 0.1%(体积比) | 0.5 mL |
Fig. 1 Proportion of nuclei in nuclear suspension obtained by different sample preparation methods A: Density map of low temperature steel-bullet beating. B: Density map of liquid nitrogen grinding method. C: Density map of blade cutting method. D: Proportion map of nucleus. The significance of the difference was analyzed by t test. **** indicates P<0.001; n =30
Fig. 2 Histograms of nuclear DNA content in nuclear suspension obtained by different sample preparation methods A: Low temperature steel-bullet beating method. B: Liquid nitrogen grinding method. C: Blade chopping method. D: M4 peak values of three sample preparation methods. E: M6 peak values of three sample preparation methods. F: M4M6 peak value between three sample preparation methods. The significance of the difference was analyzed by t test. ns indicates no statistically significant difference. n=30
制样方法 Sample preparation method | 1个样品耗时 Time taken for 1 sample/min | 30个样品耗时Time taken for 30 samples/min |
---|---|---|
低温钢珠打样法Low-temperature steel-bullet beating method | 0.5-1 | 1-2 |
液氮研磨法 Liquid nitrogen grinding method | 2-3 | 60-90 |
刀片切碎法 Blade shredding method | 6-7 | 180-210 |
Table 2 Comparison of sample preparation time of three sample preparation methods
制样方法 Sample preparation method | 1个样品耗时 Time taken for 1 sample/min | 30个样品耗时Time taken for 30 samples/min |
---|---|---|
低温钢珠打样法Low-temperature steel-bullet beating method | 0.5-1 | 1-2 |
液氮研磨法 Liquid nitrogen grinding method | 2-3 | 60-90 |
刀片切碎法 Blade shredding method | 6-7 | 180-210 |
Fig. 3 Density and histogram of tetraploid potato HTJ349-3 and diploid potato IVP101 A: Density map of diploid IVP101. B: Density map of tetraploid HTJ349-3. C: Histogram of diploid IVP101. D: Histogram of tetraploid HTJ349-3
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