Ahmad, Husaini

Hairul Azman, Roslan

Md Anowar, Hossain

Ngieng, Ngui Sing

Unimas Institutional Repository

Current Applied Science and Technology Vol. 20 No. 2 (May-August 2020) 208  Sago Palm Genome Size Estimation via Real-Time Quantitative PCR  Hairul Azman Roslan1*, Md Anowar Hossain2, Ngieng Ngui Sing1 and Ahmad Husaini1  1Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia 2Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi 6205, Bangladesh  Received: 9 December 2019, Revised: 23 February 2020, Accepted: 27 February 2020   Abstract  Sago palm, Metroxylon sagu Rottb., is an underutilized indigenous food crop that can be found mainly in the South East Asia and Pacific regions. It is a main starch producer and socioeconomically important crop in the South East Asia region including Malaysia. The sago starch provides for considerable potential to food security in the places where it is grown. However, not many molecular works have been reported thus far. In the post genomic era, sago plant genome sequencing is very important for sustainable starch development in these regions. Therefore, determination of the genome size is prerequisite to full genome sequencing and assembly. Here we report on the use of real-time quantitative polymerase chain reaction (qPCR) in determining the genome size. For this work, we calculated the genome size,  (bp) of M. sagu based on qPCR-derived copy number of two single copy genes. Pichia pastoris, with a known genome size, was used as a control to estimate sago palm genome size. With this technique, the genome size of M. sagu was calculated to be 1.87 Gbp. This genome size information would be beneficial for subsequent molecular work including genome sequencing and analysis on this economically important crop plant.  Keywords: Genome size, Metroxylon sagu, real-time PCR, copy number, Pichia pastoris DOI 10.14456/cast.2020.10  1. Introduction  Sago palm (Metroxylon sagu Rottb.) is a palm that is widely distributed in the South East Asia region. It is a starch-producing crop and economically important to the state of Sarawak in Malaysia. Sarawak has the largest acreage planted with sago palm and exports sago starch to countries such as Taiwan, Japan, Singapore and others, generating incomes of up to US$10.8 million/year [1]. Despite its importance, little research in molecular biology has been completed on M. sagu molecular biology, and it has not been sequenced, because genome size is not known for this emerging crop. The genome size is a prerequisite in genome sequencing project where it is needed to calculate the number of clones needed to be generated in shotgun sequencing and library  _______________________ *Corresponding author: Tel.: +60 82 58 3038   Fax: +60 82 58 3160              E-mail: hairulroslan@hotmail.com  

Sago Palm Genome Size Estimation via Real-Time Quantitative PCR

http://ir.unimas.my/id/eprint/29309/1/Sago%20Palm%20Genome%20Size%20Estimationvia%20Real-Time%20Quantitative%20PCR_Abstract.pdf

Sago Palm Genome Size Estimation via Real-Time Quantitative PCR

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