Frequently Asked Questions

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Frequently Asked Questions
General

I have received the sequencing results, how can I process the payment?

Macrogen Europe will issue an invoice via email after data delivery. Invoices can be paid by credit card (only Visa/MasterCard) or bank transfer. EZ-Seq, Eco-Seq and prepayment invoices are issued within 1 to 3 business days after order placement.

I've placed several orders, can I receive only one invoice?

This depends on the type of service used. Please contact info@macrogen-europe.com for Sanger sequencing orders or ngs@macrogen-europe.com for NGS orders.

What are the bank account details of Macrogen Europe?

Bank transfer to the bank account of Macrogen Europe

Beneficiary: Macrogen Europe B.V.

IBAN: NL36 INGB 0008 8449 96

Bank name: ING bank

BIC: INGBNL2A

What are your payment terms?

Macrogen Europe will issue an invoice after data delivery and our payment terms are NET 30 days.

Can I change the billing information on an invoice I've received?

Please contact our sales department via info@macrogen-europe.com for Sanger sequencing invoices or ngs@macrogen-europe.com for NGS invoices.

Why is there 21% VAT added to my invoice?

Under EU law, we have to add VAT to our invoices as follows:

  • For customers in the Netherlands, 21% VAT will be added to all invoices.
  • For EU customers outside the Netherlands, 21% VAT will be only be added to invoices if you don’t have a valid VAT number.

How can I check whether my payment has been received?

If you want to confirm receipt of your payment, or if you have any inquiries/issues regarding a payment you made, please email ar.eu@macrogen-europe.com.

Are you GDPR compliant?

Yes, please check out our Privacy Policy and Data Security Policy.

What are your T&Cs?

These terms and conditions are applicable to NGS orders:  

A. Sequencing raw data and its analyzed result will be discarded 3 months after the data delivery. A specific request for long term storage is to be negotiated separately.

B. Whilst we are willing to examine client queries surrounding the data, we dictate that any formal objections should be addressed within 3 months after the data delivery.

C. Processed sample and any unused sample (back-up, QC failed one, etc.) will be discarded 3 months after the data delivery, unless otherwise special request is made.

D. At request, the samples can be returned at the recipient’s cost. Request should be made no later than 1 month after the data delivery. Usual shipping charge is $100 to $200 depending on the region. For the project amount of total $15,000 or above, the shipping and handling charge can be waived.

E. During data storage period, analysis result of your sample might be used for sample quality check process, data validation, internal education or other investigation for better service by Macrogen.

Can I work at Macrogen Europe?

The door of Macrogen Europe is always open. If you have a passion to join a global player in the field of genomics, please take a look at our Careers page – or send your CV to directly us. Also, many internship possibilities are available. Please contact us at recruit@macrogen-europe.com.

Frequently Asked Questions About
Sanger Sequencing

Introduction

What is EZ-Seq?

EZ-Seq is a DNA sequencing service developed by Macrogen Europe. It uses 2D barcode labels for simple and secure tracking of samples. It provides DNA sequence data with a high reliability and a fast turnaround time.

How does EZ-Seq work?

When you place an order, we ship the EZ-Seq labels to you via regular mail. All you need to do is to mix the template with sequencing primer you want to use and ship the samples to our laboratory with EZ-Seq labels on the samples.


By the time you make an order, we already have your email address so the EZ-Seq labels issued are automatially linked to your email address. The sequencing data from the samples with the labels will be sent straight to your email address. This means that you do not need to log in or fill in an order form everytime you want to sequence a sample.

What are the differences between the four EZ-Seq services?

With our EZ-Seq service you can choose between tubes and 96-well plates, and between purification and purified template. The combinations give you four services. EZ-Seq purification service will go through PCR purification before sequencing.

Order Placement & Shipping

Can I give my labels to my colleagues?

Yes, you can. But remember that whoever uses the labels, and wherever the samples with labels come from, the sequence data will be directed to the designated email address, so you will need to forward the results to your colleagues.

Can I change the recipient email address from what was given in the order?

Change of delivery email address is a change of final beneficiary, and it is similar to a change of contract, so it is our recommendation not to do this frequently. However, we are flexible in accommodating your needs so we can do that for you, but we will need to get written confirmation from both parties. Please contact us if you wish to proceed with this.

Is there any order sheet necessary?

No. You can order directly through our website, and there's no need for typing in or importing the individual reaction information.

Results

How long does it take until I receive the results?

EZ-Seq is 24-hour service. From the moment of sample arrival, it will take between 12 and 24 hours depending on when the samples arrive. However, we guarantee 24 hours if there's no problem with the sample condition.

What kind of sequence results do I get?

We provide the following files.

AB1: Raw chromatogram file with quality score
SEQ: Sequence in plain text
PHD: Quality score file
PDF: AB1 files in PDF format for image processing or printing

They are compressed in a ZIP file and the download link is sent in the result email.

What's your maximum read length?

This depends on the sample condition, but in ideal condition we read about 1,050 bases – with Q20 or higher in more than 650 bases.

Payment

I have submitted a prepayment request via the ordering site, when can I expect the invoice?

We will process the request and send the invoice to you within 1 to 2 working days.

Is there a time limit on using my prepaid credits?

There is no time limit at the moment. If this policy changes we will update you.

I want to use a prepayment from a colleague’s Macrogen account, what should I do?

Our confidentiality policy does not allow us to share a user’s prepayment account without the permission of the account owner. Please ask your colleague to contact us by email and give the authorization.

Why isn’t my barcode invoice deducted from the prepayment?

Sanger sequencing prepayments are normally only used for non-barcode sequencing services. If you would like to use it for barcode invoices as well, please notify us.

Sample condition

I want to use one of the universal primers. Can I get this primer?

When you make order, you can tell use your preferred primer. We will send this primer along with your labels.

Can EZ-seq work on large-sized molecules like BAC or gDNA?

The success rate of sequencing reaction is in proportional to the number of molecules, not the amount of DNA. So even when we have same amout (ng or ug) of DNA, the number of molecules will be highly variable depending on the size of the DNA, that is, the smaller, the better. So generally the success rate of large molecules like BAC of gDNA is much lower than PCR products of plasmid. But by increasing the concentration of template DNA, we can match up the number of molecules to a certain degree. When you want to sequence these large molecules, we advise you to use the DNA with highest concentration possible.

What is the optimal concentration of DNA and primer?

In all cases, the volume of template and primer is 5ul + 5ul to make total of 10ul. The concentration varies depending on the services.

EZ-seq (without purification)
Template : 50~100ng / ul
Primer : 5pmole / ul

EZ-seq Purification
PCR product : 50ng/ul
Primer : 10pmole / ul

I'm in trouble!

I lost my labels.

Don’t worry. Your usage of label is kept in our LIMS(Laboratory Management Information System) server. Let us know of your situation and we will reissue the labels and send them to you. These labels will have same serial number with your previous labels, so it is important to discard the old labels even though you find them later as our LIMS validates every label by the time it is scanned.

I didn’t get the results yet.

There are two possibilities.

  1. Failure of sample delivery: Please provide us with the tracking number of your samples.
  2. Failure of sample processing: Due to some reasons (such as lid is open on the way, or no DNA in the tube) the samples may not be processed to save your labels. In this case, our staff may have contacted you via email. If you didn’t receive this email even in SPAM folder, please contact us immediately. (info@macrogen-europe.com)

Miscellaneous

How can you make the sequencing so inexpensive?

As many people know, the largest portion of sequencing cost is the dye terminator, machinery depreciation, and labor cost. Among these, cost saving in dye terminator already reached its saturation point, so the price comes from the other two. Thanks to the early adoption of our high-throughput sequencers, we already finished depreciation of the machines. Also by automation in the laboratory, the labor cost got much lowered. More than anything else, we are a scientists-dedicated company, and we keep our price to the level that we think is reasonable regardless of the market price.

Frequently Asked Questions About
Next Generation Sequencing

Introduction

What kind of NGS technology do you provide?

On our NGS services page you can find information on all our services and a comparison table.

What is the overall workflow of an NGS project?

Please check the NGS workflow document on our Support page.

Order Placement & Shipping

How should I pack my samples?

Please check the packing guidelines on our Support page.

How can I ship my NGS samples to Macrogen?

Contact your Macrogen Europe representative for detailed shipping instructions and to get access to our online shipment system. For information on preparing and packing samples, see our Support section.

Sample Submission

How should I prepare my DNA samples?

Start by checking the requested input amount from the QC criteria on our Support page.

  • Quantify your sample using a fluorescence-based method
  • Check the quality of your sample by gel electrophoresis
  • Aliquot your DNA samples in nuclease-free water or TE buffer in clearly labeled 1.5–2.0 mL microcentrifugetube sealed tightly with parafilm

Include a brief description of the extraction and/or enrichment protocol used in the “Comments” field of the order sheet.

How should I prepare my RNA samples?

Start by checking the requested input amount from our QC criteria.

RNA:

  • Quantify your sample using the fluorescent-based method.
  • Check quality of your sample by BioAnalyzer or equivalent.
  • · The optimal condition for RNA storage and/or delivery is ethanol precipitation method (stable for 1 year at -20C) Alternatively, RNA transport reagents from various commercial companies can be used. e.g. Ethanol Precipitation
    1. All solutions should be nuclease-free or DEPC-treated water.
    2. Add 0.1 volume of 3 M Sodium Acetate (NaOAc, pH5.5) to RNA solution and mix gently.
    3. Add 2 volume of 100% ethanol to RNA solution and mix gently. If you have 100㎕ of RNA solution, add 10㎕ of 3 M Sodium Acetate (NaOAc, pH5.5) and 220㎕ of 100% ethanol to RNA solution and mix well gently.
  • Aliquot your RNA sample in nuclease free water in a clearly labeled 1.5~2.0ml microcentrifuge tube sealed with parafilm tightly. Refer to our packing guideline. If dry ice shipping is needed, please contact your Macrogen representative.

Include a brief description of extraction and/or enrichment protocol used in the “Comments” fields of the order sheet.

Quality Checks

Which method does Macrogen use for DNA QC?

Quantity of DNA

Macrogen quantifies the starting genomic material by a fluorescence-based quantification*, rather than a UV-spectrometer-based method.

This is because fluorescence-based methods, which employ a double-stranded DNA specific dye, will specifically and accurately quantitate dsDNA even in the presence of many common contaminants. UV spectrometer methods based on 260 OD readings are prone to overestimating the DNA concentration due to the presence of RNA and other contaminants commonly found in gDNA preparations.

* Picogreen (Invitrogen, cat.#P7589)

Purity of DNA

Absorbance measurements at 260 nm are commonly used to quantify DNA.

The ratio of absorbance at 260nm to absorbance at 280nm is used as an indication of sample purity, and values of 1.8-2.0 are considered indicative of relatively pure DNA.

Condition of DNA

Gel electrophoresis condition: 1% agarose gel, 30min running at 160V, 0.5ul of DNA loaded, with 1kb marker

Size checking of DNA [Upon request, Extra fee will be charged]

  • DNA fragments <1kb : 2100 Bioanalyzer** is used for checking the size.
    **Macrogen use DNA 1000 chip and DNA 7500 chip for normal PCR product, high sensitivity chip for very small amount of DNA fragment such as ChIPed DNA.
  • DNA fragments < 150kb : PFGE method is used for large size of DNA

Which method does Macrogen use for RNA QC?

Macrogen quantifies RNA samples by a fluorescence-based quantification. We also check their integrity with an RNA Integrity Number (RIN) value using an Agilent Technologies 2100 Bioanalyzer or equivalent.

RNA that has DNA contamination will result in an underestimation of the amount of RNA used. We recommend including a DNase step with the RNA isolation method. However, contaminant DNA would be removed during mRNA purification.

It is very important to use high-quality RNA as the starting material. Use of degraded RNA can result in low yield, over-representation of the 5' ends of the RNA molecules, or failure of the protocol.

Which method does Macrogen use for Illumina library QC?

To verify the size of final library fragments, we check the template size distribution by running on an Agilent Technologies 2100 Bioanalyzer using a DNA 1000 chip.

In order to achieve the highest quality of data on Illumina sequencing platforms, it is important to create optimum cluster densities across every lane of every flow cell. This requires accurate quantitation of DNA library templates. So, we quantify prepared libraries using qPCR according to the Illumina qPCR quantification protocol guide.

Data Analysis Result

Can I see a sample of your bioinformatics analysis report?

Please contact your Macrogen Europe representative for an example report of your chosen application or Contact us here.

How can I check the integrity of my downloaded files?

You can use the MD5 checksum to verify the integrity of downloaded files. When you download your result files from the secure FTP links, do not decompress them straight away, but check the md5sum values of the files first and compare them with the provided md5sum in the report. If the values of md5sum are correct, this means that there is no forgery, modification or omission.