**Function introduction**

1）T test was used to analyze the difference between the two groups of samples.The t-test is to use the t-distribution theory to infer the probability of differences，and to compare whether the difference between the two averages is significant.

2）Wilcoxon rank sum test was used to analyze the difference between the two groups of samples.The Wilcoxon rank sum test, also known as the Mann-Whitney U test, is a nonparametric test.Whether there is a significant difference in the distribution between test samples without the assumption that both sample spaces are normally distributed.

3）Add a significantly different line to the box plot between two samples.

**Input files**

(1) Difference test form file: Enter the form file, the first column is the name of the different comparison group, each subsequent lineis the data for each iteration of the comparison group.

(2) Repeat at least three times.

(3) Pairwise comparison of multiple samples

**Parameter**

(1) X-axis title: Enter the abscissa name.(The name can only be in English)

(2) Y-axis title: Enter the ordinate name.(The name can only be in English)

(3) Chart title: Enter the entire chart name.(The name can only be in English)

(4) Statistical test method selection: T test or wilcox test.

**Results**

Default output excel result table, box plot

Ttest.Pvalue. xls：T test result (parameter selection T test, output is T test result table).

wilcox.Pvalue. xls：Wilcoxon rank sum test result (parameter selection wilcox test, output is Wilcoxon rank sum test result table).

T-test.diff.pdf/png：T test box plot (parameter selection T test, output is T test box plot).

wilcox.diff.pdf/png：Wilcoxon box plot (parameter selection wilcox test, output isWilcoxon box plot).

**1.Input file:**

Input table file, must be TXT format.Optionally, open the data in excel and save it as "text file (TAB separated)(*.txt)".

Sample form:

A |
1.35 |
1.69 |
7.2 |
8.2 |
1.3 |

B |
3.57 |
4.28 |
6.91 |
7.2 |
3.68 |

C |
22 |
12.5 |
17.4 |
22.1 |
22.9 |

**Difference test form**

2. Result output:

**
**

compare | p.format | p.signif | method |

A-vs-B | 0.52188 | ns | T-test |

A-vs-C | 0.00034 | *** | T-test |

B-vs-C | 0.00091 | *** | T-test |

3. output graphics

3. output graphics

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Note: The abscissa is the name of the two comparison groups, and the ordinate is distribution value of sample data. * represents significance,* represents two groups difference less than 0.05 ，** represents two groups difference less than 0.01，*** represents two groups difference less than 0.001，and so on.