DNA Sequence Analysis
Instrument: ABI 310
ABI 310 is a DNA analysis device based on capillary electrophoresis. This device automates the gel pouring and sample loading processes. Sequence analysis and DNA fragment analysis have become quite easy with the fluorescent color detection system.
With the 61 cm silica capillary, up to 650 bases of DNA can be read, and ten samples can be examined per day. Up to 450-500 bases of DNA can be read with a 45 cm capillary, and the processing time is shortened by approximately 1 hour per sample compared to a 61 cm capillary.
Fluorescent Dye Marking Strategy
DNA sequencing is done using the BigDye Dideoxy Terminators system.
5' fluorescent-labeled primers should be used for DNA fragment analysis (microsatellite and AFLP).
Multi-Color Detection
ABI 310 Genetic Analyzer can detect five fluorescent colors simultaneously in the same capillary. Due to this feature, the sequence analysis reaction is carried out in a single reaction tube. In genotype determination, the multi-color detection system provides the opportunity to simultaneously examine more than one PCR product of various sizes marked with different colors. During the preparation of each capillary-loaded sample, the fluorescently labeled size standard is added to ensure that the results of determining the length of the PCR products are consistent.
Sample Requirements
The quality of the data depends on the amount and purification of the DNA used in sample production. For details on the delivery of samples, please read the explanations below.
IMPORTANT EXPLANATIONS ON ANALYSIS REQUESTS FOR ABI 310 DNA ANALYZER
Persons who will order DNA sequence analysis and/or DNA fragment analysis (Microsatellite or AFLP) should carefully read the following explanations.
Sequence Analysis
- For sequence analysis, a DNA sample or PCR product of the region to be sequenced must be provided.
- The DNA sample should be at a concentration of at least 100 ng/ml and a volume of 20 ml. The picture should be taken from the DNA sample by running it in 2 ml of agarose gel, and the picture should be submitted with the sample.
- The PCR product should be 25 ml (if purified) - 50 ml (if not purified). The concentration of the PCR product should be 80-100 ng/ml. 3 ml PCR sample should be run along with the DNA size marker on the agarose gel, and the picture should be taken, and the picture should be submitted with the sample.
- Primers specific to the DNA region to be sequenced analysis should be provided at least ten mM and 10 ml. If "internal primers" are required to be used during the sequencing, they should also be provided.
- Purification of PCR products is necessary. The purification should be done with Qiagen or Q-Biogene kits. If the PCR product comes without purification, purification will be done at an additional cost.
- If the DNA of the sample is provided for sequence analysis, an additional PCR fee is charged.
- If the DNA samples are delivered without adjusting the concentration, checking on agarose gel, and taking pictures, additional fees are charged for these procedures.
- The length of the region to be analyzed for sequence analysis should be specified. "Internal primers" must also be used during sequence PCR to obtain results in regions longer than 500 bp.
Microsatellite and AFLP Analysis
- PCR should be performed using 5'-FAM, TET or HEX fluorescently labeled primers.
- DNA with appropriately sized bands in 5 µl of agarose gel from PCR products should be checked with a ladder, and its picture should be taken. The picture must be brought at the time of the analysis request.
Laboratories
Laboratories
Instrument: ABI 310
ABI 310 is a DNA analysis device based on capillary electrophoresis. This device automates the gel pouring and sample loading processes. Sequence analysis and DNA fragment analysis have become quite easy with the fluorescent color detection system.
With the 61 cm silica capillary, up to 650 bases of DNA can be read, and ten samples can be examined per day. Up to 450-500 bases of DNA can be read with a 45 cm capillary, and the processing time is shortened by approximately 1 hour per sample compared to a 61 cm capillary.
Fluorescent Dye Marking Strategy
DNA sequencing is done using the BigDye Dideoxy Terminators system.
5' fluorescent-labeled primers should be used for DNA fragment analysis (microsatellite and AFLP).
Multi-Color Detection
ABI 310 Genetic Analyzer can detect five fluorescent colors simultaneously in the same capillary. Due to this feature, the sequence analysis reaction is carried out in a single reaction tube. In genotype determination, the multi-color detection system provides the opportunity to simultaneously examine more than one PCR product of various sizes marked with different colors. During the preparation of each capillary-loaded sample, the fluorescently labeled size standard is added to ensure that the results of determining the length of the PCR products are consistent.
Sample Requirements
The quality of the data depends on the amount and purification of the DNA used in sample production. For details on the delivery of samples, please read the explanations below.
IMPORTANT EXPLANATIONS ON ANALYSIS REQUESTS FOR ABI 310 DNA ANALYZER
Persons who will order DNA sequence analysis and/or DNA fragment analysis (Microsatellite or AFLP) should carefully read the following explanations.
Sequence Analysis
- For sequence analysis, a DNA sample or PCR product of the region to be sequenced must be provided.
- The DNA sample should be at a concentration of at least 100 ng/ml and a volume of 20 ml. The picture should be taken from the DNA sample by running it in 2 ml of agarose gel, and the picture should be submitted with the sample.
- The PCR product should be 25 ml (if purified) - 50 ml (if not purified). The concentration of the PCR product should be 80-100 ng/ml. 3 ml PCR sample should be run along with the DNA size marker on the agarose gel, and the picture should be taken, and the picture should be submitted with the sample.
- Primers specific to the DNA region to be sequenced analysis should be provided at least ten mM and 10 ml. If "internal primers" are required to be used during the sequencing, they should also be provided.
- Purification of PCR products is necessary. The purification should be done with Qiagen or Q-Biogene kits. If the PCR product comes without purification, purification will be done at an additional cost.
- If the DNA of the sample is provided for sequence analysis, an additional PCR fee is charged.
- If the DNA samples are delivered without adjusting the concentration, checking on agarose gel, and taking pictures, additional fees are charged for these procedures.
- The length of the region to be analyzed for sequence analysis should be specified. "Internal primers" must also be used during sequence PCR to obtain results in regions longer than 500 bp.
Microsatellite and AFLP Analysis
- PCR should be performed using 5'-FAM, TET or HEX fluorescently labeled primers.
- DNA with appropriately sized bands in 5 µl of agarose gel from PCR products should be checked with a ladder, and its picture should be taken. The picture must be brought at the time of the analysis request.