Sweetpotato Knowledge Portal
International Potato Center (CIP) in collaboration with the Kenya Plant Health Inspectorate Service (KEPHIS) Nairobi, Kenya, conducted a one-week training on the use of high throughput sequencing and assembly of small RNAs (sRSA) for viral diagnostics. The training was conducted 19 to 30 September, 2019 at the KEPHIS Plant Quarantine and Bio-safety station at Muguga, Kenya, with 9 participants from CIP-Nairobi (3) and KEPHIS (6), representing 6 females and 3 males. The training was conducted by Segundo Fuentes, Senior Research Associate, Crop and Systems Sciences Division, Disease Detection, CIP Lima.
The purpose of the training was to help KEPHIS to start implementing sRSA in sweetpotato for identification and diagnostics of viruses. For this, training on sRSA (a combination of wet-lab and bio-informatics) was given to key staff. During this training, participants selected their own samples (18 from sweetpotato and 8 from cassava) to prepare bar-coded libraries, following the protocol according to the daily program. Libraries were mix-pooled and taken to CIP-HQ to sequence and then analyze and identify the viruses present in them.
. Sample extraction – preparing materials and reagents
Sample extraction – macerating sample for purifying RNA
Adapter ligation: given instructions and tips how to prepare small RNA libraries
Preparing bar-coded libraries. PCR enrichment (Dr. Segundo Fuentes on the right)
A post-training exam showed a success rate of 96% (23 out of 24) in small RNA library preparation, it also indicated participants had largely grasped the concepts and procedures of sRSA. It was noted that more electrophoresis equipment (tank, tray, and comb) are required if the intention is to process a large number of samples in a round.
In addition to this, a meeting on harmonization and future LAMP work was held at CIP-Nairobi office on Sep. 30, with the participation of Segundo Fuentes, Kalpana Sharma, Abdulwahab Abdurahman, and Bramwel Wanjala.
This training was supported by the SASHA II project.
