Hey All I have data frame with 5 Samples A,B,C,D,E.

A is parent (reference)sample and rest of samples are from patients. each row represents a miRNA and value against that row in each column represents Back ground subtraction values of that miRNA in each sample. I want to perform ANOVA test in R. I am bit confused how I should perform either with Parent and one patient sample (A&B) A&C and so on). Secondly most of the ANOVA tests which I saw on google and youtube they have for example one column with data second column with different groups for the value for example

```
Weight Loss     Diet
1.2              A
22.3             A
5.4              C
33.5             B  etc
                                          A        B         C         D
hsa-miR-199a-3p, hsa-miR-199b-3p         NA 13.13892  5.533703  25.67405
hsa-miR-365a-3p, hsa-miR-365b-3p   15.70536 52.86558 18.467540 223.51424
hsa-miR-3689a-5p, hsa-miR-3689b-5p       NA 21.41597  5.964772        NA
hsa-miR-3689b-3p, hsa-miR-3689c     9.58696 44.56490 10.102051  13.26785
hsa-miR-4520a-5p, hsa-miR-4520b-5p 18.06865 28.06991        NA        NA
hsa-miR-516b-3p, hsa-miR-516a-3p         NA 10.77471  8.039662        NA
                                          E      
hsa-miR-199a-3p, hsa-miR-199b-3p         NA
hsa-miR-365a-3p, hsa-miR-365b-3p   31.93503
hsa-miR-3689a-5p, hsa-miR-3689b-5p 24.26073
hsa-miR-3689b-3p, hsa-miR-3689c          NA
hsa-miR-4520a-5p, hsa-miR-4520b-5p       NA
hsa-miR-516b-3p, hsa-miR-516a-3p         NA
```

How I should do for my data

Thanks in Advance

Best  
Adnan
To transform your data into mirna/group/value format use `melt` from `reshape` package. After performing ANOVA you can do a post-hoc T-test using `pairwise.t.test` function, use for example `p.adjust.method="holm"` for multiple testing correction. The ANOVA will tell you if group/value are dependent, while post-hoc T-test for group A versus others will tell you which groups are different from parent.