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Frequently Asked Questions (FAQ)



General LGTC questions
Who can I speak to when I have a problem?
Please send an email to our general address info@lgtc.nl and we will try to reply ASAP.



Sanger sequence questions
In what kind of concentration do I need to deliver my sample?
Please refer to our sanger sequencing protocol for questions about DNA and Primer concentration.

I haven't received my results yet?
If you didn't received any notice than please check your LIMS order account to see your order status.
If there is no .zip file ready after 5 days than please contact the LGTC.

Why are my results bad?
If you can't find the answer to the problem and you have tried multiple times (different setups) than please contact the LGTC or see this website for troubleshooting.

Where can I place a sequence order?
Use the LIMS system to place an order.



Next generation sequencing
What platform can I use?
Each platform has its benefits and disadvantages. Please contact the LGTC to make an appointment for further options.

Quality/quantity of DNA/RNA sample?
Always check your samples, for example on a Lab-on-a-Chip, because if rubbish goes in, rubbish will come out.



Bio-informatics
What hardware do I need to store and/or analyze my data?
NGS data requires a lot of disk space. The raw data you will receive can be easily up to 50 Gigabytes per lane of HiSeq data. This does not include analysis files which, depending on your experiment, can double this disk space easily.

At this moment, for a small experiment, 1 Terabyte USB disks which fit in your pocket are available for under 100 euros at most multimedia stores.

For purchasing analysis hardware, it's probably better to make an appointment with us because many aspects of a project require different solutions.

Can I use hardware at the LGTC?
There is a computer cluster that can be used for analysis. Over 100 cores and nearly 1TB of RAM can be deployed. For details please contact the LGTC.

What is the difference between reads or tags?
In most cases there is none. Both terms are used to identify sequencing products. Some people refer to reads as being unaligned and tags being aligned reads. Also see our terminology page.

What about indexing and bar-coding?
These terms are also used likewise to refer to an identifying piece of artificial sequence which can be used to extract different samples from a mixed dataset.

What is the difference between genome builds and which is best?
Which is best depends on your project. Generally, the latest build is considered most complete. At the LGTC we advise to use the latest available build. More information can be found at the NCBI

What are those strange chromosomes included in the human genome containing 'un' and 'hap' in their name?
The first are random contigs whose exact location on the chromosome is not certain. Those are thus included separately. The second are different haplotypes, these are included as alternative alleles.

How should I interpret coverage or depth?
Unfortunately, for these terms there are no clear definitions. Most likely when 20X, 20 times coverage, or a depth of 20 is reported you can expect on average 20 reads covering each position in the reference.

Sometimes a coverage percentage and a depth is reported. This likely indicates how many bases of the complete genome are covered with a minimum number of reads given as the depth.

What is SAM/BAM?
The Sequence Alignment/Map (SAM) format is a format to store alignment information. There is also a binary format (BAM) available which requires less disk space and allows faster access. See the project homepage for more information.

Are there any other resources worth reading? / I can't find my answer here!
A large community of researchers specialized in NGS have a communication platform called Seq Anwers.

Why can't I open my file on my Windows computer?
(Raw) NGS files are big. If you open such a file in Word or a spreadsheet, Windows will most likely freeze or crash. If you want to analyze the data yourself we recommend to install Linux. Besides, most free software used for NGS data analysis is especially developed for Linux.



Real-time PCR
Where can I book my time on the machine?
You can book your time using the Yahoo calendar for the Fluidigm. For the LC480 there is a agenda on top of the machine to book your time.

I have problems with my PCR, can the LGTC help?
If you cannot find the answer yourself, it is best that you contact the LGTC (info@lgtc.nl).



Lab-on-a-Chip
Can the LGTC do a Lab-on-a-Chip for me?
No, the LGTC won't provide any service for LOC (unless it is taken up in a quotation). We will give a short introduction if you are a new client to use the LOC machine. Please contact the LGTC (info@lgtc.nl) if you are a new user.

Where can I book my time on the machine?
The LGTC makes use of the Yahoo calendar so that user can book on-line, the passwords are here.

It has been a long time since I last used the lab-on-a-chip, can I get help?
Yes, if you think you are not able operate the LOC anymore, please contact Arnoud Schmitz and he will help you.

The chip failed, what can I do?
Please come to LGTC to discuss the problem so we can help you further.




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