Oct 15, 2020

Public workspaceBGISEQ-500 Sequencing V.2

Version 1 is forked from BGISEQ-500 Sequencing
This protocol is a draft, published without a DOI.
  • Jie Huang1,
  • Xinming Liang2,
  • Yuankai Xuan3,
  • Chunyu Geng2,
  • Yuxiang Li2,
  • Haorong Lu2,
  • Shoufang Qu1,
  • Xianglin Mei3,
  • Hongbo Chen1,
  • Ting Yu1,
  • Nan Sun1,
  • Junhua Rao2,
  • Jiahao Wang4,
  • Wenwei Zhang2,
  • Ying Chen2,
  • Sha Liao2,
  • Hui Jiang2,
  • Xin Liu2,
  • Zhaopeng Yang1,
  • Feng Mu2,
  • Shangxian Gao1
  • 1National Institutes for Food and Drug Control (NIFDC);
  • 2BGI-Shenzhen;
  • 3State Food and Drug Administration;
  • 4BGI-Qingdao
  • GigaScience Press
  • BGI
Hongling Zhou
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External link: https://doi.org/10.1093/gigascience/giy144
Protocol Citation: Jie Huang, Xinming Liang, Yuankai Xuan, Chunyu Geng, Yuxiang Li, Haorong Lu, Shoufang Qu, Xianglin Mei, Hongbo Chen, Ting Yu, Nan Sun, Junhua Rao, Jiahao Wang, Wenwei Zhang, Ying Chen, Sha Liao, Hui Jiang, Xin Liu, Zhaopeng Yang, Feng Mu, Shangxian Gao 2020. BGISEQ-500 Sequencing. protocols.io https://protocols.io/view/bgiseq-500-sequencing-bngzmbx6Version created by Hongling Zhou
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 15, 2020
Last Modified: October 15, 2020
Protocol Integer ID: 43257
Abstract
 BGISEQ-500 is a new desktop sequencer developed by BGI. Using DNA nanoball and combinational probe anchor synthesis developed from Complete Genomics™ sequencing technologies, it generates short reads at a large scale. 
Test the quality of the sequencing library (see protocol for library preparation) by the Qubit® ssDNA Assay Kit and homogenized at 6ng total amounts.
Carry out Rolling circle amplification（RCA）for 10 minutes in an 80 ul reaction volume with pure water, buffer and DNB polymerase
Duration00:10:00
Add 20 ul DNBs stopping buffer to stop the RCA reaction
Check the quality of the DNBs using the Qubit® ssDNA Assay Kit , concentration should be above 10 ng/µL


Add 33ul DNBs loading buffer to the DNBs product from the last step, and place it in the BGIDL-50 (the DNBs loading machine)
install the sequencing chip and selected the DNBs loading process (Version: sample load 2.0) to load the DNBs.


after loading, take the sequencing chip out and install in the BGISEQ-500 （DNBSEQ-G50）sequencing machine
load the reagent sequencing kit and open the sequence control software (Version 1.1.0.10003), and select the sequence process Version 1.0.06 and Zebracall process Version 0.5.0.13875 for sequencing.
Sequencing is initiated after the sequencing reagents preloaded and sequencing chip was installed, and this process is finished in about 72 hours. When the whole sequencing was finished, the binary file with bases and quality score were converted into FASTQ format with Phred+33 quality score.