Small RNA Isolation from T. gallinae

Total RNA of T. gallinae was prepared with Trizol reagent according to the manufacturer’s protocol (Invitrogen Co. Ltd). Small RNA was prepared as described previously [20 (link)]. Briefly, small RNAs of 20–35 bases in length were isolated from 10 μg total RNA using a 15% TBE-Urea polyacrylamide gel. After adding the 5’ and 3’ adaptors (Illumina Co. Ltd), the fragments were reverse transcribed and then purified with 6% TBE PAGE gel. All gels and kits were purchased from Invitrogen Co. Ltd.

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Xu M.J., Qiu S.B., Nisbet A.J., Fu J.H., Shao C.C, & Zhu X.Q. (2014). Global characterization of microRNAs in Trichomonas gallinae. Parasites & Vectors, 7, 99.

Publication 2014

Trizol reagent 5’ and 3’ adaptors

Gels Polyacrylamide gel Trizol Urea

Corresponding Organization :

Other organizations : Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Moredun Research Institute, South China Agricultural University, Yangzhou University

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Variable analysis

independent variables

Preparation of total RNA of T. gallinae with Trizol reagent
Isolation of small RNAs of 20-35 bases in length from 10 μg total RNA using a 15% TBE-Urea polyacrylamide gel
Addition of 5' and 3' adaptors (Illumina Co. Ltd) to the isolated small RNA fragments
Reverse transcription of the fragments
Purification of the fragments with 6% TBE PAGE gel

dependent variables

Not explicitly mentioned

control variables

Manufacturer's protocol (Invitrogen Co. Ltd) for Trizol reagent preparation of total RNA
Previously described method [20] for small RNA preparation
Use of gels and kits from Invitrogen Co. Ltd

controls

No positive or negative controls were explicitly mentioned in the provided information.

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