Technology Overview
The Core has equipped with two automated nucleic acids extraction systems to offer consistent sample processing with minimized potenital variations introduced by manual labour. They allow DNA and/or RNA extraction from various sample types.
Nucleic acids are isolated from lysates in one step through binding to the silica surface of magnetic particles in the presence of a chaotropic salt. The particles are then separated from the lysates with magnetic force. After washing, pure DNA or RNA is eluted.
Purified nucleic acids are ready for use in downstream applications e.g. MassArray genotyping, Sanger DNA sequencing and Next Generation Sequencing.
QIAsymphony
It is a high-throughput nucleic acids extraction system.
Dedicated kits and customized protocols for different sample types are available on request.
| Kit | Sample Type |
| QIAsymphony DSP DNA Kit | Blood and blood-derived samples, tissue, cell cultures |
| QIAsymphony DNA Investigator Kit | Forensic and human identity samples |
| QIAsymphony DSP Virus/Pathogen Kit | Plasma, serum, CSF |
| QIAsymphony DSP Circulating Kit | Plasma, urine |
| QIAsymphony DSP AXpH DNA Kit | Liquid-based cytology samples |
| QIAsymphony RNA Kit | Tissue, cell culture |
| QIAsymphony PAXgene Blood RNA Kit | Human whole blood in PAXgene Blood RNA Tubes |
| QIAsymphony mericon Bacteria Kit | Enriched culture |
| QIAsymphony Certal Kits | Bioprocess purification buffer, cell culture supernatant samples, vaccine preparations |
QIAcube
QIAcube is a low throughput nucleic acid extraction system for processing QIAGEN spin columns. Up to 12 samples can be processed per run and no change of purification chemistry is required.
Most of the spin-column kits can be processed in QIAcube. RNA extraction kits and Allprep kits for DNA/RNA extraction are available. Please enquire for customized projects and protocols.
| Kit | Service Type |
| RNeasy Mini | Animal tissue and cells |
| miRNeasy Mini | Animal tissues and cells – Aqueous phase |
| AllPrep DNA/RNA FFPE Kit | FFPE tissue sections |
| AllPrep DNA/RNA Mini | Animal tissues and cells |
| Allprep DNA/RNA/miRNA Universal | Frozen tissue, OCT frozen tissue and Cells |
Contact us for advice on which instrument best fits your need.
Service Charges and Ordering
Service Charges
Effective 1-Sep-2023
| Service | Sample Type | Sample Input[2] | Full Service[3] | |
| HKU Academic Research | Other Academic Research | |||
| QIAsymphony DNA Extraction[1] | Buffy Coat | 220 µl | HK$ 150 | Please enquire |
| 420 µl | HK$ 165 | |||
| Whole Blood | 420 µl | HK$ 165 | ||
| 1050 µl | HK$ 185 | |||
| Serum / Plasma | Please enquire | |||
| Stool | ||||
| Saliva | ||||
| Bacteria Culture / Lysate | ||||
| QIAcube Connect DNA and RNA Extraction | OCT Frozen Tissue Sections | Please enquire | HK$ 650 | Please enquire |
| Cells | HK$ 650 | |||
| Frozen Tissue | Please enquire | |||
| QIAcube Connect RNA Extraction | Frozen Tissue, Cells | Please enquire | ||
| Other Extraction Services | FFPE, others | Please enquire |
Sample tube label[/caption]
QIAsymphony elution tube label
QIAcube elution tube label
Due to safety concerns and variations in sample type, pre-treatment (i.e. manual lysis) for samples is generally not included in above services. Please discuss with us if you have special needs.
For commercial clients, please enquire.
*IMPORTANT:* To qualify for HKU pricing, the investigator must be a regular employee of HKU AND payment must be made from an internal HKU financial account that qualifies for internal transfer. Overhead charge is required for all incoming external funding. Service Ordering Please submit service request through iLab.
Sample Collection and Storage
Tissue
- For most tissue types, a sample size of 10-30 mg is recommended.
- For heart tissue, up to 20mg and for muscle tissue, up to 40 mg may be used.
- Store tissue samples at -80°C.
Whole Blood
- Whole blood samples treated with EDTA, ACD, or heparin as anticoagulant can be used.
- Use EDTA if high molecular-weight DNA is required.
- Store blood samples at -80°C.
- To avoid repeated freezing and thawing, contact us to obtain designated sample tubes before sample collection.
Buffy Coat
- Required volume of buffy coat depends on the enrichment made during buffy coat preparation.
- Store buffy coat samples at -80°C.
- To avoid repeated freezing and thawing, contact us to obtain designated sample tubes before sample collection.
Other sample types
- Please enquire.
Workflow
Sample Submission
* Samples should be submitted within 3 months after collection of sample tubes.
* Maximum of 2 separate sample submissions are included per order of extraction service. An extra HK$ 300 handling fee is required for each additional submission.
- Submit service request through iLab.
- Contact platform specialist to arrange the collection of sample tubes, lysis buffer and digestive enzyme (if applicable).
Note: For QIAsymphony, 2 mL tubes with cat. no 72.693.005, 72.694.005 and 72.694.006 from Sarstedt should be used.Falcon 14 mL tubes with cat. no 352051 and 352057 can be used for sample volume >2000 μl. For QIAcube, 1.5 mL Eppendorf DNA LoBind tubes, PCR Clean with cat. no. 0030108051 or 2 mL tubes with cat. no 72.694.406 from Sarstedt should be used. Please enquire for compatibility of other tube types. Any incompatible tube will NOT be accepted as they will cause extraction failure due to mis-fitting on the instrument.
- Label sample tube clearly on the side with sample ID and date. Barcode labels for sample tubes will be provided if sample ID list is sent to us in advance.
- Pre-treat and transfer samples/lysates into designated sample tubes following the guideline for QIAsymphony or QIAcube extraction. Please contact us for the guidelines.
- For OCT frozen section extraction using QIAcube, the sections should be collected in 1.5 mL Eppendorf DNA LoBind tubes (cat. no. 0030108051). Please enquire for number of sections and its dimensions as optimizations/trials may be required.
- Screw tube caps tightly.
- Place the tubes in box with lid. Keep tubes in upright position.
- Download sample submission form according to the sample type and extraction method. Information in this file is used to print tube labels and these are provided at no extra cost.
Extraction using QIAsymphony Blood, buffy coat, tissue lysate, buccal swab lysate, serum, plasma, CSF, Urine: Extraction_Sample_Submission_Form_General Bacteria culture: Extraction_Sample_Submission_Form_Bacteria Extraction using QIAcube OCT frozen tissue: Extraction_Sample_Submission_Form_QIAcube frozen tissue Cell lysate: Extraction_Sample_Submission_Form_QIAcube cell line Frozen tissue: Extraction_Sample_Submission_Form_QIAcube frozen tissue Other sample types: Extraction_Sample_Submission_Form_General - Enter sample IDs into the worksheet “sample information”. Attach the form under service request in iLab. Print a hardcopy.
Sample ID should be of maximum 10 characters.
“Space” and special characters e.g. ~_!@#$%^&*() are NOT allowed. - Contact platform specialist to arrange sample submission.
- Bring sample tubes in box to the core. Samples should be kept on ice or dry ice (depending on the sample type) during transportation.
Nucleic Acids and Data Collection
Nucleic Acids
DNA will be eluted in RNase-free water, Buffer AVE, ATE or EB with minimum elution volume available. For other elution volumes, please enquire.
Store DNA at -20°C or -80°C to avoid degradation.
RNA will be eluted in RNase-free water with minimum elution volume available. For other elution volumes, please enquire.
Store RNA at -80°C to avoid degradation.
User is responsible for collecting the nucleic acids from our centre within 3 months after completion of the job.
DNA Information
For Full Service (except ccfDNA extraction), quantity and quality of extracted DNA will be assessed by NanoDrop and agarose gel electrophoresis respectively.
NanoDrop data and gel images will be emailed to user with notification of DNA collection.
In order to eliminate the absorbance reading contributed by the magnetic beads, A320 should be subtracted for accurate assessment of concentration.
DNA Concentration = 50ug/mL x (A260-A320)
Corrected A260/A280 = (A260-A320)/(A280-A320)
Corrected A260/A230 = (A260-A320)/(A230-A320)
Secure your own copy of gel photos and NanoDrop data. The Core will not be keeping the data indefinitely.
FastRuler High Range DNA ladder is run beside your samples for size reference.
RNA Information
For full service of RNA extraction, quantity and quality of extracted RNA will be assessed by NanoDrop, Qubit and Bioanalyzer.
Data Delivery
NanoDrop data, Qubit results and gel images will be included in an extraction result file. Bioanalyzer results will be sent in PDF format. All the extraction results will be sent to user via ODDS with notification of nucleic acid collection.
Secure your own copy of result files. Genomics Core will not be keeping the data indefinitely.
Processing Time
Time for completion of service will depend on the number of samples submitted and the level of service demand at sample submission.
Your job will be put in processing queue as soon as all samples and required information are submitted to the Core.
Delay may be due to unforeseeable circumstances and long queue for service, in which the user will be notified at the time of submission.
Terms of Service
Please kindly inform if sample contains known infectious agent.
We will be responsible for the proper operation of instrument.
The quality or quantity of extracted DNA cannot be guaranteed as both are dependent on the original sample quality. Actual elution volume also varies depends on samples.
Quality of extracted DNA (except ccfDNA) will be assessed by agarose gel electrophoresis and quantified by NanoDrop.
The Core has successfully used the extracted DNA for downstream work, e.g. Sequenom genotyping and NGS.
Carryover of trace amount of magnetic beads into the DNA samples is expected. User may spin down the beads and use the supernatant for downstream experiments.
Invoice will be sent in the following month after completion of job.
Contact
Mr TSEA, Ivan
Ms LAU, Tracy
