Introduction
Core technology
Facilities
Instruction
Business hours
Charge
Lab members
Certification
Forms
Related links
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Instruction
- According to standard operation procedure of testing and instrument operation for capillary automatic DNA sequencer, Genome Sequencing Center, Testing
Laboratory, Department of Medical Research and Development (MRD-SOP-SC-001).
- The main facility : ABI 3730 DNA Analyzer (capillary automatic DNA sequencer).
- Please provide Plasmid, PCR products or other samples to be sequenced. And adjust the concentration of the samples and volume to the following table:
DNA type |
DNA concentration
(ng / £gl) |
DNA volume (£gl) |
PCR DNA (<1000 bp) |
> 20 |
> 10 |
PCR DNA (>1000 bp) |
> 40 |
> 10 |
Plasmid DNA |
> 200 |
> 10 |
- Please fill out two forms :
- DNA sequencing application form (MRD-SOP-SC-001-F02) (with DNA Template electrophoresis photo, and indicate the loading
volume and the concentration of Marker)
- Test commissioned form (MRD-POC-003-F01)
- Provide a variety of samples, please note:
- Customer must provide DNA information (such as: GC, GT, or GCGT rich) and the concentration of appropriate purified DNA samples
- Customer must provide appropriate primer concentration (> 5 £gM), or use standing primer (M13F, M13R, T7P, SP6, T3)
- Quality of DNA Template is very important to sequencing results, so customers must provide high quality and "clear " Template, excluding the extra protein,
RNA and other impurities. Purified DNA samples OD260/280 should be between 1.7 and 1.9, OD200/260 should be around 1.1. Lower OD260/280 means that samples
contain higher level of protein content, higher OD260/280 means that higher level of RNA samples and genomic DNA; higher OD200/260 means that samples
contain genomic DNA and Salts.
- Sequencing primer purified by PAGE is better
- Concentration:> 5 £gM.
- No mispair.
- Only one binding site with template.
- No hairpin structure, especially in the 3 '-end.
- 18-24 bp in length , Tm = 55 ~ 60 ¢J.
- 3'- end should be end in GC, the percentage of GC between 50 to 55% is ideal.
- Sequencing primer should avoid four or more than four repeat sequence.
- If you deliver the samples after 3:30 pm, that will be considered as next working day cases.
- You could get your reports within seven working days at the collection counter. (The first working day is the next day of receiving the samples).
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