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Sequencing | Sample Submission Guide

To submit your project and ensure its smooth and speedy completion, please follow these 5 steps:

1. Check that you have sufficient amount and quality of sample:

Please ensure that your samples meet the general sample requirements listed below, and that you can provide the amount, concentration and volume of nucleic acid for the specific service you require. This can be found on page 2 of your quote or follow the links for the application specific submission criteria below. If you are unable to provide the amount or quality of material required please contact us.

General Sample Requirements:

  • Samples must be intact and not degraded.
  • Samples must be clean, 260:280 and 260:230 ratios >1.8.
  • Lower 260:230 ratios are acceptable if a column purification method was used.
  • RNA samples must be free of genomic DNA contamination.
  • DNA samples must be free of RNA contamination.
  • Samples must be resuspended in nuclease free water and not TE – PacBio submissions excepted. 

Application Specific Sample Requirements:

 

2. Arrange for payment: 

  • Send us a copy of your purchase order. We regret that we are unable to initiate projects without a copy of a purchase order.
  • If you would like to pay by credit card, request an invoice from us and provide proof of payment prior to submitting samples. 
  • If you are from UNSW, include your UNSW project code on the electronic submission form. 

3. Submit your project online:

  • Projects must be lodged in our Laboratory Information System (LIMS) before shipping. The system will issue a unique LIMS ID for each sample submitted. 
  • If you are a returning customer you can log in to our LIMS system here.  
  • New customers are required to sign up for a user ID. Refer to our guide to submitting samples for detailed information on the online submission process. 

4. Prepare your samples for shipping:

  • Projects with <24 samples can be submitted in 1.5 ml microfuge tubes.
  • Do not use screw cap or strip tubes, or seal tubes with parafilm, foil or tape. 
  • Projects with >24 samples must be submitted in 96-well PCR plates. 
  • All submissions must be clearly labeled with their unique LIMS ID.
  • Tubes must be labelled with individual LIMS IDs and plates with the project LIMS ID and plate number. 
  • If one of our technical specialist has advised you of the requirement to batch your samples, ensure that they are randomised and in the correct order for batch processing. 

5. Ship your samples:

  • Our recommended couriers are World Courier and PDP Couriers. 
  • Ensure plates are well sealed. We recommend using a foil seal overlaid with plastic seal to prevent puncture.
  • Wrap plates or tubes in bubble wrap and place in a sealed plastic bag.
  • Use granular dry ice. Do not use dry ice blocks. 
  • For domestic shipping please ensure there enough dry ice to last for two days shipping. 
  • If you are shipping from overseas ensure the courier will top up the dry ice as required and has the documentation for Australian customs e.g. AQIS import permit
  • Include a printed copy of the electronic submission form with the package, this helps identify your samples.
  • Notify us by email when your samples have been shipped and provide us with an approximate delivery date.

 

Courier Delivery Address
Ramaciotti Centre for Genomics
Upper Campus Store, Room LG018
E26, Bioscience South
LG018 Loading Dock,
Via Gate 11 Botany Street,
UNSW Sydney NSW 2052
Australia
Tel: (02) 9385 1241
Hours: Monday to Friday, 8.00am – 4.00pm

In Person Drop off
Ramaciotti Centre for Genomics
Level 2, Biosciences South Building (E26)
Botany Street
UNSW Sydney, NSW  2052
Tel: (02) 9385 1241
Hours: Monday to Friday, 8.30am – 5.00pm
Please check that there will be someone available to accept them by calling (02) 9385 1241.

 

 

Application Specific Sample Submission Criteria

Genomic DNA Sequencing

  • Consult the table below for the amount, concentration and volume of DNA required for each assay. Please also ensure that samples also meet the general submission criteria.
  • DNA must be double-stranded and not degraded as assessed by 0.8% agarose gel. Please include gel images when shipping your samples.
  • DNA concentration must be measured using the Qubit or Picogreen assay. Quantification by NanoDrop is inaccurate and should not be used for quantification. If this is not possible in your laboratory please contact us. 

Flip device horizontal to see full table

SERVICEAMOUNT REQUIREDCONCENTRATIONMINIMUM VOLUME
DNA Seq (PCR-Free)3 ug (350bp insert)
5 ug (550bp insert)
50 ng/ul
60 ng/ul
60 ul
80 ul
DNA Seq Nano (PCR)300 ng (350bp insert)
500 ng (550bp insert)
10 ng/ul
15 ng/ul
30 ul
30 ul
Nextera DNA400 ng20 ng/ul20 ul
Nextera XT DNA100 ng5 ng/ul20 ul

 

RNA-sequencing

  • Consult the table below for the amount, concentration and volume of RNA required for each assay. Please also ensure that samples also meet the general submission criteria. 
  • For submission of rRNA depleted RNA please verify rRNA depletion by Bioanalyzer and provide traces pre- and post- depletion with your submission.
  • For rRNA depletion Illumina recommends Epicenter’s RiboZero kit or Ambion’s MICROBExpress kit.

Flip device horizontal to see full table

APPLICATIONINPUT & AMOUNT REQUIREDCONCENTRATIONMINIMUM VOLUME
mRNA Seq 3 ug total RNA70 ng/ul40 ul
Total RNA Seq - rRNA depleted submission300 ng rRNA depleted RNA10 ng/ul25 ul
Total RNA Seq RNA - human, mouse, rat 3 ug total RNA*100 ng/ul30 ul
Total RNA Seq RNA - bacteria or epidemiology3 ug total RNA*100 ng/ul30 ul
small RNA3 ug total RNA200 ng/ul15 ul

*If submitting samples for a low input assay please contact the Centre.

ChIP-Seq, MeDIP-Seq and Nucleosomal Sequencing

  • 10ng of the enriched immunoprecipitated material in a total volume of 20ul.
  • The quantity and quality of the input material should be checked by a fluorescence assay (Qubit or Picogreen) and by the quantitative real time PCR, respectively.
  • Please supply us with the fragment size range of the enriched material (200-600bp).
  • Where possible the size range of the immunoprecipitated material should be assessed by Bioanalyzer using the High Sensitivity kit.
  • IMPORTANT – It is not recommended to use salmon sperm DNA as blocking agent during the immunoprecipitation as it could contribute to contamination of your sequencing data.

Amplicon/Microbial Community Profiling

  • 15-20ul of DNA at a concentration of 5-10ng/ul (based on Qubit/PicoGreen quantification). 
  • DNA of high purity (260/280, 260/230 >~1.8).  
  • Samples should be submitted in 96-well PCR plates in consecutive columns with no gaps between samples.
  •  94 samples is the maximum number that can be submitted per plate. Two wells are required for positive and negative controls.
  • Samples must be amplifiable by PCR. Please check this prior to submission and provide a gel photo. 
  • If your samples require any special conditions or additives (i.e. BSA, DMSO) to amplify, please let us know at the time of submission.

Libraries (User Supplied)

  • >25ul of library at a concentration of 4nM (based on qPCR and/or Qubit/PicoGreen quantification, and Labchip/Tapestation/Bioanalyzer for sizing).
  • Library fragments up to 900nt.
  • If custom primers are required these must be supplied at the time of sample submission.
  •  Only one pool per run can be submitted, unless prior arrangements have been made.

Long & Linked Read Sequencing

Please download our Long & Linked Read Sample Submission Guide for further information on DNA/RNA quality and concentration. 

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