How tos and FAQs

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Which qPCR instruments do you provide training for?

We can train you regardless of qPCR instrument you have in your lab. For a long time we focus on instruments by Applied Biosystems ...

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You offer refurbished sequencers, what exactly does that mean?

These are instruments that were already at use and their previous owners decided not to use them anymore for various reasons, typically not sufficient throughput or on the other hand infrequent usage. In our hands these machines are refurbished. This means they are disassembled piece by piece,..

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I am interested in qPCR courses but there are no dates announced.

We hold private courses, exclusively at the client's request.

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I want to register for your workshop as a private person, how can I do this?

Our registration form is primarily designed for clients whose participation at courses is covered by their employer and therefore a standard invoice address with valid VAT is required during registration. In case you are a private person and have no VAT number please get in touch with us and we shall process your registration outside of our website. Thank you for your understanding.

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I got lost which sequencer to choose, please advice!

The key decision maker is the throughput. This means how many samples you analyze at the same time.

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How do I order Next-Generation Sequencing?

All orders are placed through our web interface. To order analyzes, you must...

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How do I order Sanger sequencing or Fragment analysis?

All orders are placed through our web interface. To order analyzes, you must...

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Why should I choose SEQme as an instrument or service provider?

We have long experience with servicing and technical support for DNA sequencers and Real-Time PCR instruments.

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General rules of sample preparation

When ordering sequencing or fragment analysis in tubes, preferably please use 0.2 ml strips to send samples, they are easy to work with.

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Sample submission guidelines

Please always follow steps described in our Sample submission guidelines.

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What are the electrical requirements when installing sequencers 3130 or 3130xl?

We recommend using a UPS - Uninterruptable Power Supply ...

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Is it necessary to air-condition the room where the sequencer is installed?

The recommended temperature range for the working site is 15-25°C.

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How do I ship samples?

We recommend you insert samples into a quick-closing plastic bag. Fragment analysis samples can be (not necessarily) wrapped up in aluminium foil to prevent light access.

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Guidelines for Ready-to-Run libraries (Illumina only)

Here are our requirements for accepting libraries prepared by our clients.

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Guidelines for sample preparation for DNA metabarcoding - Tailed-amplicon sequencing (Illumina only)

Samples must be purified after first PCR and delivered exactly in required concentration

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Sample submission guidelines - Oxford Nanopore sequencing

Please read these recommendations before submitting samples for Oxford Nanopore sequencing.

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StandardSeq – Preparation of samples

Sample is a template mixed together with sequencing primer in a single test tube / well or template only if you wish to use some of our universal primers. Template is a PCR product or plasmid.

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Sample shipment guidelines

It depends what samples you have and how they are processed, but it is generally advisable to send samples cooled...

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RapidSeq - Preparation of samples

Please use the instructions for the StandardSeq protocol.

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ColonySeq – Preparation of samples

Sample is bacterial cells from agar plate colony, liquid media culture, or glycerol stock. This protocol is designed for mid- to high-copy number plasmids (> 10 copies per cell).

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CleanSeq – Preparation of samples

Sample is an unpurified PCR product (aliquot of the PCR reaction) or other suitable template, amenable to enzymatic cleanup. You must also supply sequencing primer(s) or select some of our universal primers.

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HairpinSeq – Preparation of samples

Sample is typically a plasmid with a cloned hairpin or GC-rich stretch in the region of interest or PCR product. Price of the HairpinSeq protocol includes bidirectional sequencing (forward and reverse).

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What is the difference between sequencing protocols you offer?

StandardSeq is the basic protocol, we sequence your template. CleanSeq is in fact StandardSeq with template cleanup performed beforehand. HairpinSeq is a special protocol ...

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How are RapidSeq protocol results evaluated?

The evaluation of results obtained by the RapidSeq protocol takes place without the intervention of human hands.

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Fragment analysis – Preparation of samples

The sample is typically a PCR product or a mixture of PCR products which are fluorescently labelled. The PCR products do not have to be purified, you only need to send the aliquot of the PCR reaction.

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Can I copy-paste into the order form?

Definitely yes. Create (in Excel, for example) a table with three columns - Sample Name, Primer Sequence and Template Size (applies to StandardSeq, CleanSeq, ReadySeq). For HairpinSeq the table must have four columns - Sample Name, Primer1 Sequence, Primer2 Sequence and Template Size.

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How do I measure DNA/RNA concentration?

Please measure DNA/RNA concentration with a fluorescence-based measurement such as Qubit or Picogreen assay. These measurements tend to be specific for DNA/RNA.

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Universal primers

The following universal primers are supplied by us free of charge.

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My own primers

When designing custom primers, please remember that properly designed primer is crucial for sequencing...

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What target regions do you use for taxonomic analysis?

Various hypervariable gene regions or whole genes can be used for sequencing taxonomic analysis.

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How should I specify primer sequences?

Please use ACGTN symbols for the primer sequences, IUPAC codes (BDHRYKMSWV) for degenerate bases and “I” for Inosine. If there are...

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I want to use my own primer which you use as a universal primer, how can I do this?

If you want to use for example your own M13F primer, you have two choices. Either you do not enter primer information in the order form and only add it to your template (StandardSeq protocol), or you specify this primer in My Primers...

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How should I specify the template size?

If the template is a PCR product, enter its size. If the template is plasmid with a cloned insert, enter the total size of the template (plasmid plus insert). This field is optional for ReadySeq.

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There is a limit on template length in the order form for Sanger sequencing. Why?

This is correct, the template size must be between 100 bp and 20.000 bp. It is quite unusual to sequence templates shorter than 100 bp. Assuming your primer has 20 bases ...

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Can I copy-paste into the NGS order form?

Definitely yes. Create (in Excel, for example) a table with three columns: Sample Name, Concentration, Volume. The sample names must be unique...

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Should I provide any specific data about my samples when submitting NGS order?

Yes. When submitting an order you will be asked to specify concentration and volume of each sample you send ...

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What sequencing capacity should I allocate?

This does depend on many factors, namely number of samples you submit, average fragment length...

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I cannot send the online order

If, after entering data into the order form and clicking on the Order button, you get the notification that no sample data has been entered

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What data will I receive after my sequencing experiment has been finished?

You can receive data (results) in any format you wish. However, if you have no specific requirements, you will receive from us the following standard sequencing outputs.

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Can you guarantee what amount of NGS sequencing data I will get?

It depends on the type and course of order processing.

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Where and how long do you store my results?

All Sanger/Fragment analysis orders are processed on our website. Results will be made accessible in your user profile and can be downloaded from within.

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What happens with my NGS samples in SEQme lab?

The first step is a quality control (concentration measurement, gel visualization, bioanalyzer etc.).

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What if my NGS samples do not pass quality control in SEQme lab?

The key to a successful NGS experiment is starting with a sufficient quantity of high quality material.

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Will the data I finally get be analyzed?

This does depend on what you order. All samples always undergo a primary data analysis ...

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Where and how long do you store my NGS results?

All NGS orders are processed on our website. Results will be made accessible in your user profile, including but not limited to raw data, analysis results, supplementary data etc. and can be downloaded from within.

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Subscribing sequencing services

Click on link New Prepayment in your account and complete the form to subscribe to our services. The amount to be prepaid is given without VAT, the system will add VAT automatically according to applicable regulations. Once you have confirmed your order, an advance payment request will be issued automatically which you can use to ...

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Subscribing sequencing services

Click on link New Prepayment in your account and complete the form to subscribe to our services. The amount to be prepaid is given without VAT, the system will add VAT automatically according to applicable regulations. Once you have confirmed your order, an advance payment request will be issued automatically which you can use to ...

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Other questions?

Do not hesitate to contact us
We will be glad to help you.

Richard Nádvorník
+420 602 102 790
info@seqme.eu

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