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Welcome to ChunLab Support
ChunLab FAQ
1. What do I need to do to prepare my sample(s)?
For microbial community analysis, you need to submit extracted metagenomic DNA or PCR amplicon products produced by your lab.

• Submission of DNA samples: the amount of DNA submitted must be sufficient to perform quality check and several amplification reactions. DNA samples should have a volume that is at least over 30 ul and have a concentration that is over 30 ng/ul. At this level, we will be able to successfully conduct the experiment, even if PCR fails. It is always better to send a higher amount of DNA to ChunLab, as any leftover DNA may be returned after the community analysis, if requested. If the DNA concentration and volume are insufficient, we will have no choice but to return the DNA.
• Submission of amplicon samples: amplicon samples are PCR products in which amplification using barcoded primers have already been performed at your lab. To perform pyrosequencing on these amplicons, we need to know the barcode sequences and the concentration of the PCR product. To properly conduct pyrosequencing, the PCR product should have a concentration that is over 50 ng/ul and a volume that is over 30 ul when measured by a nanodrop.
2. What kind of extraction method or kit do you recommend?
There are various extraction kits provided by different companies. You can use any extraction kit that is ideal for the extraction of DNA from your particular sample. Please ensure that you provide us with a high enough concentration and amount of DNA for the community analysis. But please keep in mind that you should use the same extraction kit to extract DNA from all the samples used in a comparative analysis. We use a DNA extraction kit for soil (provided by MP Bio) to extract most environmental genomic DNAs.
3. How do I ship samples to ChunLab?
We recommend sending DNA or amplicon samples packed in an ice box in order to ensure the reliability of the data. This is especially true for environmental samples as there is a possibility that the microbial community could undergo changes during shipment. To ensure the analysis is performed on the original community, you must send environmental samples refrigerated. If an ice box is not used for the shipment of DNA or amplicon samples, an express shipment method must be utilized. For more information, please contact us at chunlab@chunlab.com. Shipments sent to ChunLab should be addressed as follows:

4. Can I use different PCR conditions for each sample?
You can use different PCR conditions for your samples. However, all PCR conditions should be identical with regards to the target environmental sample for comparative analysis of a microbial community. Differences found in a community could arise due to the extraction method used or the PCR amplification condition set. Please note that these differences are not a result of differences inherent in the community's origin, but are caused by different experimental processes used. We especially recommend the inclusion of a humic acid removal step for soil samples, as humic acids greatly inhibit PCR. Applying this humic removal process to all DNAs will lessen the possibility of PCR amplification failure, and will improve the efficiency of the experiment. Once more, we emphasize that all experiments should be conducted under the same conditions for proper comparative analysis of samples.
5. How long does it take to get the results of a community analysis?
Results vary heavily depending on the type of sample submitted (i.e.: DNA or amplicon). While we typically provide a complete analysis of a community within a maximum of 45 to 60 days, after the samples pass DNA extraction quality check, certain samples may need more time due to different experimental processes.
6. How much does it cost?
Cost varies according to the type of sample submitted. For a specific quotation, please send an email to chunlab@chunlab.com, indicating the type of sample and the number of samples you plan to submit. Once we receive your information, a detailed quote will be provided as soon as possible.
7. What is the submission procedure for samples?
The submission process is presented in our Sample Preparation Guideline for Microbial Community Analysis. First, contact us to receive a quote. Second, submit your sample order sheet. Third, register at our homepage at www.chunlab.com. All your pyrosequencing results and the CLcommunityTM software (bioinformatics tool for analyzing your results) can be downloaded on our homepage. When your samples arrive at ChunLab, our experiment team will perform the preconditions for pyrosequencing then our analytic team will analyze the sequences. Shortly after we finalize our analysis of your data we will contact and you can then download the results from our website.
8. How can I make fusion primers?
• A fusion primer is a barcoded primer for community analysis. Detailed information about fusion primers can be found on http://www.ezbiocloud.net/resource/method/content?uid=M1001.
• What should be considered for the submission of amplicon samples?
If you conducted PCR amplification using fusion primers, each sample should contain a unique barcode as we distinguish a sample’s origin with the barcode. The size of the amplicon should be approximately 500~600 bp; and all short fragments, long fragments and primer dimmers must be depleted during purification.
9. How do I use the CLcommunityTM software for analysis?
The CLcommunity™ User Manual and CLcommunity™ User Tutorial can be downloaded from www.chunlab.com. In addition, a help menu is available within the CLcommunity™ software itself. Furthermore, we hold regular workshops and webinars focused on how to do analyses within CLcommunity™ as well as other experiment protocols. It would be helpful to participate in one of our workshops (www.chunlab.com). If you are interested in registering for a webinar, contact us at chunlab@chunlab.com.

©2014-2015 ChunLab, Inc.
Data generated by ChunLab is for research purposes only. It is not intended for diagnostic use.
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