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Sample preparation guide

NGS

Sample preparation guide


De Novo Sequencing of Animal and Plant Genomes


Total Sample Amount: For small fragment library preparation, a total amount greater than 1 μg of sample is required. For mate-pair libraries (2 kb, 5 kb, 10 kb), a total amount of 20-50 μg is needed. To ensure experimental quality and continuity, please provide at least 2 times the sample preparation volume at once.


Sample Concentration and Purity: Sample concentration should be >200 ng/μl, with an OD260/280 ratio between 1.8 and 2.0, and no visible protein, RNA, or other impurities.


Sample Quality: The genome should be intact without degradation. The main band of genomic DNA on electrophoresis should be above 23 KB after Hind III digest and clear without smearing.


Sample Preservation: Please store samples in dry powder, alcohol, TE buffer, or ultrapure water, and indicate this on the sample information sheet.



Re-sequencing of Animal and Plant Genomes


DNA Sample Amount: Prepare 1 μg of sample for each library construction, and it's recommended to provide 2 times the preparation volume for single sample submission.


Sample Concentration and Purity: Sample concentration should be >50 ng/μl, with an OD260/280 ratio between 1.8 and 2.0, and no visible impurities. The main band in electrophoresis should be greater than 23 kb.


Sample Selection: For plant samples, select dark-grown seedlings or young seedlings. For animal samples, choose tissues with low fat content, such as muscle or blood.


Sample Preservation: Store samples in dry powder, alcohol, TE buffer, or ultrapure water.



Whole Exome and Targeted Region Re-sequencing


DNA Sample Amount: Please provide at least 1 μg of high-quality genomic DNA.


Sample Concentration and Purity: Sample concentration should be >50 ng/μl, with an OD260/280 ratio between 1.8 and 2.0, and no visible contamination.


Sample Quality: The genome should be intact without degradation. The main band of genomic DNA on electrophoresis should be greater than 23 kb.


Sample Selection: Same as for the previous category.


Sample Preservation: Same as for the previous category.



De Novo Sequencing of Microbial Genomes


DNA Sample Amount: For small fragment library preparation, a total amount of 3 μg of sample is required. For mate-pair libraries (2 kb, 5 kb, 10 kb), 10-50 μg of sample is needed. To ensure experimental quality and continuity, please provide at least 2 times the sample preparation volume at once.


Sample Concentration and Purity: Sample concentration should be >100 ng/μl, with an OD260/280 ratio between 1.8 and 2.0, and no visible contamination.


Sample Quality: The genome should be intact without degradation.


Sample Selection: For bacteria and fungi samples, try to obtain them from the same strain, and avoid host DNA contamination.


Sample Preservation: Store samples in dry powder, alcohol, TE buffer, or ultrapure water.



Transcriptome Sequencing


RNA Sample Amount: ≥ 1.0 μg of total RNA is required.


Sample Concentration and Purity: Sample concentration should be >100 ng/μl, with an OD260/280 ratio between 1.8 and 2.0, and no visible contamination. For animal RNA, RIN >7 is required, and for plant RNA, RIN >6.5 is required.


Sample Selection: Same as for the previous category.


Sample Preservation: Store samples in ethanol, DEPC water, and indicate this on the sample information sheet.


Sample Transport: Place samples in 1.5 ml tubes, seal them with sealing film, and transport them on dry ice.


The remaining categories follow similar requirements for sample preparation, concentration, purity, quality, selection, preservation, and transport.




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Guangzhou IGE Biotechnology Co., Ltd.
Email: marketing@igebio.com
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Address: Building G1-903, South China New Materials Innovation Park, #31 Kefeng Road, Huangpu Dist. Guangzhou, Guangdong prov. China 510663

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