Materials and Reagents

1. Filter papers
   
2. Plastic pots
   
3. Young Vigna mungo plants
   
4. Whiteflies (Bemisia tabaci)
   
5. HgCl₂
   
6. TruSeq small RNA Library Preparation Kit (Illumina, catalog number: RS-200-0012 )
   
7. Trizol reagent (Invitrogen, USA)
   
8. Isopropanol
   
9. Acrylamide/bisacrylamide (Merck Millipore, catalog number: 623100281001730 )
   
10. T4 RNA ligase 2, truncated (New England Biolabs, catalog number: M0242 )
    
11. Tris-HCl pH 7.5
    
12. MgCl₂
    
13. DTT
    
14. ATP
    
15. PEG8000
    
16. SuperScript^® II reverse transcriptase (Thermo Fisher Scientific, Invitrogen^TM, catalog number: 18064014 )
    
17. TEMED (Merck Millipore, catalog number: 623171280051730 )
    
18. Ammonium persulphate (Merck Millipore, catalog number: 623171100101730 )
    
19. 10x TBE buffer (Thermo Fisher Scientific, Ambion^TM, catalog number: AM9865 )
    
20. UltraPure^TM glycogen (Thermo Fisher Scientific, Invitrogen^TM, catalog number: 10814-010 )
    
21. 3 M sodium acetate (Sigma-Aldrich, catalog number: S7899-100ml )
    
22. Absolute ethanol (EMD Millipore, catalog number: 100983 )
    
23. Qubit^® dsDNA HS Assay Kit (Thermo Fisher Scientific, Invitrogen^TM, catalog number: Q32854 )
    
24. High Sensitivity DNA Kit (Agilent Technologies, catalog number: 5067-4626 )
    
25. T4 RNA Ligase 2 buffer (see Recipes)
    

Equipment

1. Electrophoresis apparatus (Bio-Rad Laboratories, model: Mini-Sub^® GT Cell )
   
2. Bioanalyzer (Agilent Technologies, model: Agilent 2100 )
   
3. Thermocycler, DNA engine cycler (Bio-Rad Laboratories, model: PTC-0200G )
   
4. Qubit 3.0 fluorometer (Life Technologies)
   
5. Illumina genome analyzer IIx (Illumina)
   

Software

1. Cutadapt-0.9.3
   
2. Bowtie-0.12.7
   
3. SeqQCv2.1
   
4. BLAST (Rfam and miRBase)
   

Procedure