Redefining Reference Standards

What is the impact of molecular assay failure in your laboratory?
Identify and eliminate variability in your laboratory today

FAQs

You are here: Home / Connect With Us / FAQs

Below we have compiled a list of frequently asked questions and suggested solutions. 

  • Ordering FAQ
    • How to place an order

      We accept orders using the following methods:

      Online: Our online catalog and ordering system can be used to search our full menu of available reference standards. Select your desired reference standards and add them to the shopping basket. Your order can be submitted instantly using credit card, telephone ordering or a purchase order number as the preferred payment method.

      Please click here to access our online catalog and ordering system.

      Offline: If you prefer to send us orders outside our online ordering system, please see the Contact Us page for telephone, fax, and email details in order to place an order.  We recommend that you use our online system to identify the products you require and their respective “HD” product codes. To order offline we will require the following information:

      • Product code(s) of the item(s) you wish to order (e.g. "HD123")
      • Your name, email address, title and organization
      • Shipping address and billing address with contact names
      • Purchase Order Number if you have one
      • Telephone number
      • VAT Number (European Countries only)
    • Is the product in stock?

      All products visible in the online ordering system are in stock and available.

    • Why should you use our online ordering system?

      Speed: Our online ordering system has been designed to allow us to receive the information we need to process your order more rapidly

      Immediate confirmation: Upon receipt of the order you will receive a confirmatory email detailing your order for your records and so that you can Contact us should there be any errors 

      Fast and easy re-ordering: Your details are stored for future purchases.  Just log in and your personal information will be pre-loaded. 

      Easy access to discounts: Price plan discounts are available on your online account and will provide preferential product pricing. Please Contact us for more information on our pricing plans.

    • Do you charge VAT?

      Our prices are exclusive of VAT and VAT only applies to customers in the European Union (EU).  If you are based in the UK and are VAT exempt, a copy of your VAT exemption certificate must be provided with your order (fax or email).

  • Shipping FAQ
    • What are your shipping charges?

      The following table outlines our shipping charges.  Our online system will automatically apply the appropriate shipping cost based on order size. Shipping prices will be reviewed on an annual basis and are subject to change.

        Standard (FedEx)

      Europe

      €15.00

      North America

      $20.00

      UK

      £13.00

      ROW

      $20.00 - $100.00

    • What is the delivery time?

      We aim to ship your order within five (5) to ten (10) business days of receipt of your order and we will notify you of any changes that may affect your delivery time.

    • What support can we expect once the order is shipped?

      Shortly after shipping of your order, Horizon Diagnostics will contact you to confirm that you have received the shipment in good condition and that you are satisfied.  We will also reach out periodically thereafter to see if you need any support or have any questions.  And of course you can Contact us at any time for technical support or product inquiries.

    • What are the storage conditions of your Reference Standards?

      Stability studies support storage of HDx™ DNA, FFPE and FISH reference standards at 4°C. You can store our DNA products at -20°C but we recommend that you avoid multiple freeze-thaw cycles (>10) to avoid DNA shearing and loss. 

    • Why do you ship at Room Temperature?

      For our FFPE and DNA products, stability and transport studies indicate that our reference standards are stable for over 8 weeks at 37°C and shipping at room temperature reduces packaging wastage.  

  • HDx™ DNA Reference Standards FAQ
    • What are the storage conditions of your DNA Reference Standards?

      Stability studies support storage of HDx™ DNA Reference Standards at 4°C and are stable for at least two (2) years. You can store HDx™ DNA Reference Standards at -20°C but we recommend that you avoid multiple freeze-thaw cycles (>10) to avoid DNA shearing and loss.

    • What is the concentration and pack size of your DNA Reference Standards?

      Our singleplex HDx™ DNA Reference Standards are available in 100ng vial sizes 20μl at 5ng/μl. We do have some starter pack sizes available (20ng mutant + 200ng wild type) but these packs are being phased out. Higher concentrations are available for HDx™ multiplex DNA Reference Standards.

      If you require bulk quantities or a customised concentration of our HDx™ DNA Reference Standards please Contact Us and outline your requirements.

    • What is the difference between HDx™ DNA Reference Standards and plasmids/oligonucleotides?

      All HDx™ DNA Reference Standards are manufactured using purified human cell line DNA and contain the entire human genome. This provides a contextual reference point for your assay. They do not contain short oligonucleotides or plasmid DNA.

    • How do I generate a dilution curve with your DNA Reference Standards?

      A dilution curve of allelic frequencies can be generated using any 50% mutant HDx™ DNA Reference Standard and its matched wild type (0% mutant) genomic DNA reference standard. A suggested dilution curve is outlined below and for this you require 1x 100ng 50% mutant vial and 1x 200ng wild type pack:

      Allelic Frequency

      Mutant ( /µl)

      Wild type ( /µl)

      20%

      4

      6

      10%

      2

      8

      1%

      1µl of 10% Dilution

      9

      0

      0

      10

    • How are your DNA Reference Standards manufactured?

      All HDx™ DNA Reference Standards are produced by extracting genomic DNA from a human cell line cell pellet generated from adherent cells grown in a monolayer.

    • How do you quantify your DNA Reference Standards?

      We quantify our HDx™ DNA Reference Standards using the Quantifluor dsDNA assay (Promega) that uses a fluorescent DNA-binding dye that enables sensitive quantitation of dsDNA. In addition we monitor expected copies/μl using digital PCR for the gene of interest. 

    • Can I purchase DNA that has been extracted from your FFPE Reference Standards?

      Currently we only supply DNA extracted from our cell lines. This DNA can be used to generate stoichiometric dilution curves of defined allelic frequency.

      If you are interested in custom production of genomic DNA extracted from FFPE we can offer this as a service. Please Contact Us to discuss your requirements.

    • How do you validate and verify your DNA Reference Standards?

      HDx™ Genomic DNA, Quantitative Multiplex and FFPE Reference Standards are manufactured using our ISO 13485 certified processes. We confirm the mutation sequence using Sanger Sequencing and confirm the allelic frequency using our QX100™ Droplet Digital™ PCR System. A summary of our quality control workflow can be viewed here.

  • HDx ™ FFPE Reference Standards FAQ
    • What is the format of your FFPE Reference Standards?

      We provide our HDx™ FFPE Reference Standards as thin FFPE sections/curls (10μm or 15μm) contained within tubes. The FFPE section is NOT slide mounted. DNA extraction can be performed directly on FFPE sections contained within these tubes.

    • How do you manufacture your FFPE Reference Standards?

      Our cell lines are grown, harvested, washed and re-suspended in PBS ensuring no cell clumps are present. The cell number is precisely counted and for multiplexing the required cells from each cell line are mixed to create the defined allelic frequency. This mixture is formalin treated using proprietary conditions, pelleted and washed carefully in 70% EtOH before paraffin embedding to generate a FFPE block. All FFPE sections are prepared and analysed using our ISO 13485 certified quality control process. 

    • What are the storage conditions of your FFPE Reference Standards?

      Stability studies support storage of HDx™ FFPE Reference Standards at 4°C and are stable for at least 2 years. We do not recommend storage at -20°C due to the potential freeze-thaw damage to the paraffin wax.

    • What is the recommended method for DNA extraction from your FFPE Reference Standards?

      As part of our quality control process we extract DNA using Promega’s Maxwell LEV Plus FFPE extraction kit and achieve yields of 400ng – 700ng per FFPE sample. In addition we also extract using Qiagen’s QIAamp DNeasy kit and achieve yields of 200ng – 500ng per FFPE sample. 

    • The FFPE yield is lower than expected, how can I improve the yield?

      Using a column based method we would expect you to achieve an extraction yield of 200ng – 500ng. If you observe no yield or less than 50ng total recovered DNA it is possible that the cell pellet was lost during the xylene step and subsequent wash steps. During the xylene treatment it is important to be able to locate and visualize the clear cell pellet (a small cell disc) in your tube to ensure no cells are aspirated at this initial stage. 

      If you are experiencing total DNA yields of 50ng – 200ng per FFPE section then it is possible that some of your DNA remains on the column during the elution. Performing multiple elution steps will be beneficial. From our experience two 50μl elutions provide a greater total DNA yield compared to a single 100μl elution. 

    • How many cells are contained within each FFPE Reference Standard?

      HDx™ FFPE Reference Standards contain approximately 200,000 cells per section and a potential of 1μg total extractable DNA. Due to the impact of formalin fixation and your extraction method we would expect a DNA yield of between 200ng – 700ng from each FFPE sample.

    • Can I purchase DNA that has been extracted from your FFPE Reference Standards?

      Currently we only supply DNA extracted from our cell lines. This DNA can be used to generate stoichiometric dilution curves of defined allelic frequency.

      If you are interested in custom production of genomic DNA extracted from FFPE we can offer this as a service. Please Contact Us to discuss your requirements.

    • How do you validate and verify your FFPE Reference Standards?

      HDx™ Genomic DNA, Quantitative Multiplex and FFPE Reference Standards are manufactured using our ISO 13485 certified processes. We confirm the mutation sequence using Sanger Sequencing and confirm the allelic frequency using our QX100™ Droplet Digital™ PCR System. A summary of our quality control workflow can be viewed here.

    • Can I extract the DNA from your FFPE samples and mix to generate a custom allelic frequency?

      It is possible to mix DNA from extracted samples but it is critical that the DNA is accurately quantified using a dsDNA dye approach (e.g. Quantifluor™ or Picogreen™) and not spectrophotometry (e.g. Nanodrop), which would not be sufficiently accurate.

      For generating a dilution curve of defined allelic frequency we would recommend using HDx™ DNA Reference Standards rather than HDx™ FFPE Reference Standards. In addition we offer predefined HDx™ DNA Reference Standards validated to 50%, 20%, 10%, 5%, 1% allelic frequencies which you can use to create a standard curve without need for dilution.

  • HDx™ FISH Reference Slides FAQ
    • What is the format of your FISH Reference Slides?

      We provide our HDx™ FISH Reference Standards either as FFPE sections (5μm) OR cell spreads mounted onto microscope slides. Each reference slide will contain between 10% - 100% cells showing chromosomal abnormalities and consistent hybridization pattern within 100 scorable interphase nuclei.

    • How do you manufacture your FFPE FISH Reference Standards?

      Our cell lines are grown, harvested, washed and re-suspended in PBS ensuring no cell clumps are present. The cell number is counted and for multiplexing the required cells from each cell line are mixed to create the defined allelic frequency. This mixture is formalin treated using proprietary conditions, pelleted and washed carefully in 70% EtOH before paraffin embedding to generate an FFPE block. 5μm FFPE sections are prepared and mounted onto microscope slides.

    • How do you manufacture your Cell Spread FISH Reference Standards?

      Our cell lines are grown, harvested, washed and re-suspended in PBS ensuring no cell clumps are present. The cell number is counted, fixed using methanol:acetic acid (3:1) and spread onto a microscope slide.

    • What are the storage conditions of your FFPE FISH Reference Standards?

      Stability studies support storage of our FFPE HDx™ FISH Reference Slides at 4°C and are stable for at least 2 years. We do not recommend storage at -20°C due to the potential freeze-thaw damage to the paraffin wax.

    • What are the storage conditions of your Cell Spread FISH Reference Standards?

      Stability studies support storage of our Cell Spread HDx™ FISH Reference Slides at 4°C and are stable for at least 1 year. 

    • Can I extract DNA or RNA from your FISH Reference Slides?

      It is possible to extract DNA or RNA from our reference slides but we cannot guarantee the yield or quality of the nucleic acids recovered. 

    • How do you validate and verify your FISH Reference Standards?

      Our Cell Spread and FFPE HDx™ FISH Reference Slides are manufactured using our ISO 9001 certified process. We confirm the presence of the chromosomal abnormality using FISH analysis.

    • What FISH hybridization protocol would you recommend?

      For our FFPE HDx™ FISH Reference Slides we would recommend that you shorten the baking time and reduce pepsin concentration and incubation time to improve the quality of staining. 

      Please Contact Us for specific details.

  • HDx™ Q-Seq NGS Reference Standard FAQs
    • What is a Q-Seq NGS Reference Standard?

      We have developed HDx™ Q-Seq NGS Reference Standards that contain multiple gene mutations within one sample and these are available as either DNA or FFPE reference standards. More information can be found here

      The general quantitative multiplex reference standards contain >10 oncology focused mutations each confirmed using digital PCR. These are not titratable.

      Titratable gene specific quantitative multiplex reference standards (e.g. K-Ras or EGFR) have been developed that allow you to test for the limit of detection of your multiplex test. For example you can titrate the sample with the relevant wild type standard to create:

      Sample 1Sample 2Sample 3Sample 4
      K-RAS 16.7% G12D K-RAS 10.0% G12D K-RAS 5.0% G12D K-RAS 1.0% G12D
      K-RAS 16.7% G13D K-RAS 10.0% G13D K-RAS 5.0% G13D K-RAS 1.0% G13D
      K-RAS 16.7% A146T K-RAS 10.0% A146T K-RAS 5.0% A146T K-RAS 1.0% A146T

      More information on these can be found here.

    • How do you validate and verify your Q-Seq NGS Reference Standards?

      HDx™ Genomic DNA, Q-Seq NGS and FFPE Reference Standards are manufactured using our ISO 13485 certified processes. We confirm the mutation sequence using Sanger Sequencing and confirm the allelic frequency using our QX100™ Droplet Digital™ PCR System. A summary of our quality control workflow can be viewed here.

    • What is the concentration and pack size of your Q-Seq NGS Reference Standards?

      HDx™ Q-Seq NGS Reference Standards are available in 100ng vial sizes 20μl at 5ng/μl or 200ng vial sizes of 4μl at 50ng/μl. 

    • What is the format of your Q-Seq NGS Reference Standards?

      We provide our HDx™ FFPE Reference Standards as thin FFPE sections/curls (10μm or 15μm) contained within tubes. The FFPE section is NOT slide mounted. DNA extraction can be performed directly on FFPE sections contained within these tubes. 

    • What are the storage conditions of your Q-Seq NGS DNA Reference Standards?

      Stability studies support storage of HDx™ DNA, FFPE and FISH reference standards at 4°C. You can store our DNA products at -20°C but we recommend that you avoid multiple freeze-thaw cycles (>10) to avoid DNA shearing and loss.

    • What are the storage conditions of your Q-Seq NGS FFPE Reference Standards?

      Stability studies support storage of our HDx™ Q-Seq NGS FFPE Reference Standards at 4°C and are stable for at least 2 years. We do not recommend storage at -20°C due to the potential freeze-thaw damage to the paraffin wax.

    • What is the difference between HDx™ Reference Standards and plasmids/oligonucleotides?

      All of our HDx™ Q-Seq NGS Reference Standards are manufactured using purified human cell line DNA and contain the entire human genome. This provides a contextual reference point for your assay. They do not contain short oligonucleotides or plasmid DNA.

    • How are your Q-Seq NGS DNA Reference Standard manufactured?

      All of our HDx™ Q-Seq NGS DNA Reference Standards are produced by extracting genomic DNA from a human cell line cell pellet generated from adherent cells grown in a monolayer. More information can be found here.

    • How do you quantify your Q-Seq NGS DNA Reference Standards?

      We quantify our HDx™ Q-Seq NGS DNA Reference Standards using the Quantifluor dsDNA assay (Promega) that uses a fluorescent DNA-binding dye that enables sensitive quantitation of dsDNA. In addition we monitor expected copies/μl using digital PCR for the gene of interest. 

  • How can I contact you if I have a question?

    If you have a question regarding Horizon Diagnostics’ products or services, you can contact us in a number of ways:

    Ask Horizon a question through our website

    Contact us by phone, fax or email

  • What is the intended use of your reference standards?

    HDx™ Quantitative Reference Standards have been designed and developed to enable independent verification of your molecular assays (e.g. K-Ras, EGFR, B-Raf,...). These materials should be used routinely to monitor the accuracy of your molecular assay(s). All of our current catalogue products are for RESEARCH USE ONLY

Login Error


There was an error with your login - please try again.

Forgotten your password? Click here

OK ×

Recover your password

Please enter your email address and we'll send you a reset link

×

Thank you.


We've sent you an email to reset your password.

OK ×

Add Voucher


Sorry, Unable to add voucher

OK ×

Account Error


There was an error updating your details.

OK ×

Thank you.


Your changes have been saved.

OK ×

Thank you


Thank you for registering to the Horizon DX website. We hope you enjoy and benefit from our online resources, and please feel free to contact us if you have any queries or feedback through the website itself or at info@horizondx.com.


×

Thank you.


Items have been saved to your basket.

OK ×