3730xl DNA Analyzer Applied Biosystems Technology
User Guide
The instructions in this guide must be followed carefully to achieve the best possible results and avoid delays in processing your request.
Samples that do not comply with the recommendations may be rejected without compensation.
Note that delays in the processing of samples will vary depending on the size of the project. It is recommended to contact the Client Management Office for more information.
- Starting Material
- Requirements – Volume and concentration
- Requirements – Quality
- Solutions Accepted in Empty Wells
- Required containers – Sample Organization and identification
- Service Request Form and Sample Submission
- Sample Shipment Preparation
- Transmission of results
Starting Material
The starting material for the microsatellite marker genotyping service may vary depending on the options chosen.
| Option | Description | Starting Material |
|---|---|---|
| 1 Read only | Reading of PCR products previously resuspended in formamide with a standard molecular weight scale. | PCR product* previously resuspended in formamide + standard |
| 2 Addition of formamide with standard + reading | Reading of PCR products to be resuspended in formamide with the standard 500Liz molecular weight scale. | PCR product* |
| 3 Addition of formamide with standard + reading + analysis | Reading of PCR products to be resuspended in formamide with the standard 500Liz molecular weight scale and analysis of chromatograms. | PCR product* |
| 4 Reading + analysis | Reading of PCR products previously resuspended in formamide with a standard molecular weight scale and analysis of chromatograms. | PCR product* previously resuspended in formamide + standard |
| 5 PCR + reading + analysis (10 markers) | Carrying out PCR amplification from genomic DNA (gDNA), reading and analysis of the 10 markers.** | Genomic DNA (human only) |
| 6 PCR + reading + analysis (24 markers) | Carrying out PCR amplification from gDNA, reading and analysis of the 24 markers.** | Genomic DNA (human only) |
* All PCR products to be analyzed must have been amplified with a pair of primers, one of which is labeled with a fluorochrome. Accepted fluorochromes are FAM, VIC, NED and PET.
**For the identification of human cell lines, the kits used include the following markers:
GenePrint® 10: AMEL, CSF1PO, D13S317, D16S539, D21S11, D5S818, D7S820, TH01, TPOX, vWA.
GenePrint® 24: Amelogenin, D3S1358, D1S1656, D2S441, D10S1248, D13S317, Penta E, D16S539, D18S51, D2S1338, CSF1PO, Penta D, TH01, vWA, D21S11, D7S820, D5S818, TPOX D, YS391, D8S1179, D12S391, D19S433, FGA, D22S1045.
Requirements – Volume and concentration
| Sample type | Option | Volume (concentration) |
|---|---|---|
| PCR product in formamide | 1 and 4 | 10 µL |
| PCR product* | 2 and 3 | 10 µL |
| Genomic DNA (gDNA) | 5 and 6 | 10 µL (10 à 50 ng/µL) |
*For options 2 and 3, if the volume of 10 µL is not respected for each sample, the plate will be automatically rejected.
It is the customer’s responsibility to provide samples of good quality and sufficient quantity.
Requirements – Quality
| Sample Type | Recommended method for verifying concentration | Recommended method for verifying purity |
|---|---|---|
| gDNA | Qubit or equivalent fluorometric method. Note: Spectrophotometric methods such as Nanodrop are much less accurate for measuring DNA concentration. | Nanodrop or other spectrophotometric method. Target optical density ratios: – 260/280 between 1.8 and 2.0 – 260/230 between 2.0 and 2.2 |
| PCR Product | The concentration, size and integrity of the PCR products must have been verified by electrophoresis gel. | |
Solutions Accepted in Empty Wells
Each empty well must contain the same volume as a well that contains a PCR product by adding one of the solutions below:
- Hi-Di™ formamide
- EDTA 0.2 mM
- Ultra pure water type 1 (MilliQ water)
- UltraPure™ DNase/RNase-Free Distilled Water
Required containers – Sample Organization and identification
| Recommended tubes for gDNA | 96-well PCR plate recommended | 96-well PCR plate unaccepted |
|---|---|---|
| – PCR strip tubes of 8 or 12 All types of 200µL PCR tubes in strip of 8 or 12 with a cap are accepted. Example: 0,2 mL PCR strip tubes with 12 well, VWR, Cat # 53509-300 And strip domed caps for 12 well strips, VWR, Cat # 53509-302 | – Half-skirt PCR plate All types of clear half-skirt 96-well PCR plates are accepted. Example: Thermo-Fast 96 PCR detection plate with flat deck; Life Technologie, Cat# AB1400L Example: 96-well plate, standard semi-skirted, clear; FroggaBio, Cat#SS-96S | – Cell culture plates – No-skirt PCR plates – Full-skirt PCR plates – Opaques plates |
| – Clear adhesive film Example: Adhesive PCR Films; Thermo Fisher Scientific, Cat#AB0558 – Aluminum adhesive film Example : Adhesive PCR Foil Seals; VWR International, Cat#60941-074 – Strip caps of 8 or 12 preventing any risk of leakage and cross-contamination. |
- For the microsatellite marker genotyping service, 96-well plates can be submitted full.
- For options 1 and 4, read-only and for read-only + analysis, if a plate is not full, Type 1 ultrapure water (or another accepted solution) must be added to each empty well.
- For the human cell line identification service, two wells per plate must be left empty for the addition of controls, wells H11 and H12.
- Samples must be placed in the plate as indicated on the submission form – Row arrangement mandatory.
The name on the tube or plate must be unique, readable, concise, and identical to the name indicated on the submission form.
This is to avoid any delays in processing the request.
Service Request Form and Sample Submission
Click on service request and sample submission to view instructions.
Sample Shipment Preparation
Click on samples shipment preparation to view instructions.
Transmission of results
An email is sent as soon as the results are available.
The reading and analysis results, if applicable, are directly accessible via the Nanuq web application.
The raw data files generated by the 3730xl DNA Analyzer are in .fsa format.
Version 1.0_2025-11-06