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How can you make the sequencing so inexpensive? |
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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. |
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I didn’t get the results yet. |
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There are three 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.
3) Failure of email address : Some of institutions do not allow emails with direct link in it.
Please contact your system administrator is this is the case. If not, please contact us immediately and we will have it corrected ASAP. |
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I lost my labels. |
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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. |
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What is the optimal concentration of DNA and primer? |
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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 (without purification)
Template : 50~100ng / ul
Primer : 5pmole / ul |
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Can EZ-seq work on large-sized molecules like BAC or gDNA? |
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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. |
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I want to use one of the universal primers. Can I get this primer? |
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When you make order, you can tell use your preferred primer. We will send this primer along with your labels. |
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How do you do the purification of PCR products with primer already mixed? |
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Our R&D team successfully developed Macrogen's unique technology called MSE based on partial dephosphorylation, and could specifically eliminates the molecules with minority, and EZ-seq Purification adopts this technology. So please feel free to mix the primer. |
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Do I have free shipping of the samples? |
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For most of European countries, we keep the 20-sample free shipping policy, but there are some exceptional countries where our courier charges exceptionally high price for the shipping. In such countries, it is 30. Please contact us for details about these exceptional countries. |
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Do I have individual invoices per sample shipment? |
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No. The invoice is issued only once by the time the order for labels is made. Even if you buy 1,000 labels at a time, and send 50 samples every week, you will have only a single invoice for the 1,000 labels. So you do not need to struggle with all the paperworks. |
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What is PO(Purchase Order) number? |
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It has many different names depending on the institution you work for, but the most common name is Purchase Order number. Your institution will issue an internal reference number for every purchase made in that institution. This is what we call PO number. This number will appear on our invoice to you so that the financial manager in your institution can match the invoice with purchase. Some institutions may not have PO number, and in such case you may leave it blank. |
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How long can you read? |
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It depends on the sample condition, but in most ideal condition we read about 1,050 bases with Q20 or higher in more than 650 bases. |
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What kind of sequence results do I get? |
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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 ZIP file and the download link is sent in the result email. |
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How long does it take until I receive the results? |
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EZ-seq is 24-hour service. From the moment of sample arrival, it will take between 12 hour ~ 24 hour depending on what time point the samples arrive. But we basically guarantee 24 hours if there's no problem with the sample condition. |
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Is there any order sheet necessary? |
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No. You can make order from this web page, and there's no need for typing-in or importing the individual reaction information. You know better about your samples than we do, and we do not need to know what you are sequencing. All we do is to do our best with what we got from you! |
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Can I change the recipient email address from what was given by the time of order? |
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Change of delivery email address is the change of final beneficiary, and it is more or less like change of the contract, so it is our recommendation not to do this frequently. But we are quite a flexible in fitting ourselves to your needs, and we can do that for you. But we will need to get the written confirmation from both of two parties. Please contact us if you wish to proceed with this. |
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Can I give my labels to my colleagues? |
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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. For change of the recipient email address, please refer to the article right below. |
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Why is the email address so important? |
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In DNA sequencing service, Email address is just like delivery address of the final product. Let's say you orders one vial of Taq polymerase. The most important information in the order will be where the product should be delivered. So, having a proper and secure email address connected to the EZ-seq label is very much important. |
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Why do I have to provide all these information? |
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We tried to reduce the amount of information required, but found that there are some essential information without which we cannot properly deliver our service. So we beg your kind understanding for these. Once you've set up the account in our on-line system, you won't need to fill in those information again. Your information shall be encrypted and securely stored in our server, and shall not be used for any other purpose than correct delivery of our service. |
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How do I use the extra sample or primer which was delivered along with the previous shipment? |
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In case of such primers which are not registered in previous orders but you sent us as extra ones for the purpose of further use, it will not appear on the list of stored primers since that information does not exist in our database but it is stored manually.
In such case, you may make an Additional order and register the primer as enclosed with a comment that the primer was already sent to Macrogen. Please do not forget to write the order number of the primer. You can also send us e-mail or LIMS message to ask for additional reactions. |
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How can I save the shipping fee if I need to send samples (or primers ) repeatedly for the held reactions? |
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Please send the held ones along with your next batch including 20 or more reactions.
Otherwise, your shipping is NOT free.
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Should I check the concentration of samples before sending you? |
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To ensure proper concentration, please check your samples by electrophoresis before sending them to us. There should be a single clear band of the appropriate size when loading 2 ul of the sample on a 1% agarose gel. |
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Turn around time |
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Within 48 hours from the date of sample arrival, the results shall be sent to you.
It may take 1~2 more working days in case of primer synthesis or purification. |
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Sequencing primer |
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Ideally the sequencing primer should be designed 100 bases inside the PCR product (i.e. 100 bass downstream of the PCR binding site).
To ensure optimal results
- The length of the primer should be between 18 and 25 bases (ideally 20-25 bases)
- The desirable G-C content is 40 – 60 % with a melting temperature (Tm) around 55 -60 °C
- No significant hairpins (>3bp)
- Free of salts, EDTA, or other contaminants
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I can't place an order by online due to computer system error. |
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To avoid any delay, please fill out the order sheet which can be down loaded from the below link and send the complete order sheet to us.
- Order sheet Download |
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Is there any restriction on reaction numbers per order? |
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If there are too many reactions in one order, you may get into trouble to check the result or download the data because of overload. Up to 384 reactions are recommended for one order. So please split your order into several orders in case of over 384 reactions. |
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Can I modify the sample name (or primer name) before sequencing? |
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Yes, you can modify the reactions information while the order status is shown as Pending . However, it is unable to modify any information when the order is confirmed. |
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Do I need to place an order separately depending on the sample condition? |
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Yes, each sample should be registered separately according to its sample type as long as the conditions of experiments between PCR product and plasmid are different, so please register samples separately if their types are different. |
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How do I pack the plates to avoid potential damages in transit? |
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Do I need to place an order before sending samples to Macrogen? |
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Yes, you should summit an order first before sending samples to us.
Otherwise, the experiment may NOT be performed.
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If I have any trouble with payment, how to solve it? |
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Please contact to payment@macrogen.com |
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Which credit cards do you accept? |
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We accept Visa, Master Card, or American Express Card. |
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How do I pay for my order? |
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Once, sequencing order has been finalised the accorded invoice will be issued. Then you can choose any available payment option(Credit Card/Wire Transfer/Cheque).
More information on payment can be found from below address:
http://dna.macrogen.com/eng/support/howtopay.jsp |
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Sample / primer return |
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Macrogen does not send back DNA sample or primer to customers. Due to difficult customs clearance in some countries, Macrogen Korea cannot sometimes send out biological products.
If you have to get them back, please check below things first:
1. custom clearance in your countries and required document
2. Fedex account number of yours so that we can send as recipient's payment |
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Free Shipping to Macrogen Inc. (Korea) or Macrogen Europe (The Netherlands) |
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Macrogen Inc.(Korea) pay shipping fee for more than 20 reactions's order.
Our account number of Fedex(preferred), DHL or TNT should clearly be written and it should be checked as reciever's payment on the airway bill. We are not responsible for paying wrongly charged invoice.
It is room temperature packing only and should be less than 1kg. (Not in a big box.) If you want to pack your samples with dry ice/ice pack, shipping fee is not covered by Macrogen Inc.(Korea).
**Free shipping for European express sequencing service is also applied, so please contact Macrogen Europe at europe@macrogen.com. |
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Retrial(Re-sequencing) Request Terms&Condition |
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If you are not satisfied with the previous sequencing reactions, you can request re-trial. We will check DNA condition and primer to make a better result in the retrial. (This resequencing service is not provided for any kind of plate discount order.) The sample and primer should have the same match and both sample and primer should be available here in Macrogen. If you have to send more sample or primer, it is not regarded as a free retrial order. It will be regarded as a paid new order.
Re-sequencing request is only available for one month from the notice date of sample arrival. |
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Glycerol Concentration |
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I need to know how should I send my cells for sequencing. What percentage of Glycerol should I use in the samples.
=> 1:1 mix up with 30% glycerol stock to make 15% of final glycerol concentration would be fine. |
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Log in Problem |
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If you fails to log in even with correct ID and password, it could be cookie problems.
If browser version is 6.0, you may need to change your computer's cookies settings as described below.
1) On the Tools, select 'Internet Options'.
2) Remove cookies and all off-line files from "Delete Cookies."
3) Then, go to the "Privacy Tab" and set it "Advanced."
4) Select "Override automatic cookies handling" and "Always allow session cookies". These two options must be checked, as well as "First_Party Cookie_Accept".
5) Select "Privacy" and click "Edit".
6) Address of website "dna.macrogen.com" and select "Allow."
7) Click "OK" and shut all the exploer windows.
8) Try log in with a new explorer window. |
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Do you synthesize a primer? |
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Yes. We do synthesize a primer.
Macrogen offers the oligonucleotide DNA synthesis service applying the cutting-edge technology and the state-of-the-art facility.
We also provide free delivery for your oligonucleotide DNA through express courier service if the order is over 300 mers.
Please click the link below for more information about the oligonucleotide DNA synthesis service.
http://dna.macrogen.com/eng/support/news/news_view.jsp?board_number=5981
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Will you keep my samples? |
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We can keep leftover of DNA and primers for one month ~ three months upon receipt. If you want to sequence among them, please place an order from "Additional Order" menu. Or, you may just give us a comment when and from whom samples were delivered here with sequencing information. |
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Concentration of Primer |
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20ul with concentration of 10pmole can cover 5 reactions. Please increase 10pmole each time the number of reactions is increasing.
We recommend the quality of HPLC or PAGE purified primers for sequencing reactions, but OPC is o.k as long as it can guarantee the equivalent quality. About 17-25 mer, 40~60% of GC content and less than 4bp of dimer formation or hair-pin formation are recommended to guarantee high end stability. Tm is required about 50~60°C, because annealing temperature is fixed at 50°C. Primer concentration should be known before sequencing reactions.
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May I place an order in Maryland Branch? |
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You may also use our Maryland branch's facility. It has the same quality of sequencing skills. But it is independent from Seoul, Korea office and the price is little bit different from Korea headquarter's.
You may contact US branch and get more information.
9700 Great Seneca Hwy
Rockville MD
20850 USA
Tel : 301-251-1007
Fax : 301-251-4006
customer@macrogenusa.com |
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Is there minimum number of samples in sequencing order? |
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No, there is no such minimum number or maximum number of sequencing reactions. Just we recommend you send DNAs for at least 20 sequencing reactions to save your shipping fee. |
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What is 96 / 384 well plate sequencing? |
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If you order in 96 or 384 well plate, you may have choose either of the plate sequencing order with discount.
1. "Plate Sequencing" refers to the orders charged per plate.
To use this order, your samples should meet all of the following criteria.
- Samples are sent to Macrogen pre-arranged in 96-well plates. (No 384 plate service is available)
- Only two custom primers (or two universal primers) in tubes could be acceptable for applying this Plate sequencing.
* Note:
- For three or more primers, another paired primer plate with your sample plate should be provided.
if not, your order will be regarded as Normal sequencing reactions.
- The well to well variation of the concentration of your samples is very important to get
better results, please keep it as constant as possible.
2. "1 Primer / 1 Plate" refers to the orders charged per plate.
To use this order, your samples should meet all of the following criteria.
- Samples are sent to Macrogen pre-arranged in 96-well or 384-well plates.
- Only one single primer should be applied to the whole plate,so that we Macrogen can simply put the primers to the corresponding sample plate.
- More economical than Plate sequencing service.
- No sample name is required, but plate name is enough for this service.
We will provide the results as the following file name.
And please be aware that there is no free resequencing service in either of above discounted sequencing order. If rerun is required, it will be regarded as a new order of regular reaction.
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Do I need to use on-line system all the time? |
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On-line system is made for the ease and accuracy of order, so we recommend you to use the system. It is much more convenient because you will be able to order online and retrieve the result online as well as leave a message in no time. |
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What is the average read length? |
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We read approximately ~850bp per reaction, and 650bp of them has Phred 20 or higher quality. |
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What kind of machines you use? |
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We use ABI3730XL |
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What kind of computer system is required for on-line system? |
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The system is designed to be compatible with all of the systems including PC, Mac, and many other conventional operating system. |
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Universal primers available in Macrogen |
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| Primer Name | Sequence | Base | | Bluescript SK | CGCTCTAGAACTAGTGGATC | 20 | | EBV-RP | GTGGTTTGTCCAAACTCATC | 20 | | KAN2-FP | ACCTACAACAAAGCTCTCATCAACC | 25 | | KAN2-RP | GCAATGTAACATCAGAGATTTTGAG | 25 | | M13-FP | TGTAAAACGACGGCCAGT | 18 | | pBacPAC-RP | GTCTGTAAATCAACAACGC | 19 | | pBAD-FP | ATGCCATAGCATTTTTATCC | 20 | | pDONOR-FP | TAACGCTAGCATGGATCTC | 19 | | pEGFP_N | CCGTCCAGCTCGACCAG | 17 | | pEGFP-FP | TTTAGTGAACCGTCAGATC | 19 | | pEGFP-RP | AACAGCTCCTCGCCCTTG | 18 | | pESP-RP | TCCAAAAGAAGTCGAGTGG | 19 | | pET-24a | GGGTTATGCTAGTTATTGCTCAG | 23 | | pET-RP | CTAGTTATTGCTCAGCGG | 18 | | pMalE | TCAGACTGTCGATGAAGC | 18 | | pREP-fwd | GCTCGATACAATAAACGCC | 19 | | 35S-A | AAGGGTCTTGCGAAGGATAG | 20 | | 35S-B | AGTGGAAAAGGAAGGTGGCT | 20 | | AD Reverse | AGATGGTGCACGATGCACAG | 20 | | CYC1 Reverse | GCGTGAATGTAAGCGTGAC | 19 | | DsRed1-C | AGCTGGACATCACCTCCCACAACG | 24 | | DsRed1-N | GTACTGGAACTGGGGGGACAG | 21 | | EGFP-C | CATGGTCCTGCTGGAGTTCGTG | 22 | | EGFP-N | CGTCGCCGTCCAGCTCGACCAG | 22 | | GAL1 Forward | AATATACCTCTATACTTTAACGTC | 24 | | U-19mer Primer | GTTTTCCCAGTCACGACGT | 19 | | T7 EEV | ATGTCGTAATAACCCCGCCCCG | 22 | | Bluescript KS | TCGAGGTCGACGGTATC | 17 | | pFastBac Forward | GGATTATTCATACCGTCCCA | 20 | | pFastBac Reverse | CAAATGTGGTATGGCTGATT | 20 | | AOX1 Forward | GACTGGTTCCAATTGACAAGC | 21 | | AOX1 Reverse | GCAAATGGCATTCTGACATCC | 21 | | a-Factor | TACTATTGCCAGCATTGCTGC | 21 | | STag 18mer Primer | GAACGCCAGCACATGGAC | 18 | | MT Forward | CATCTCAGTGCAACTAAA | 18 | | QE Promoter | CCGAAAAGTGCCACCTG | 17 | | pRH Forward | CTGTCTCTATACTCCCCTATAG | 22 | | pRH Reverse | CAAAATTCAATAGTTACTATCGC | 23 | | SV40-pArev | CCTCTACAAATGTGGTATGG | 20 | | SV40-Promoter | GCCCCTAACTCCGCCCATCC | 20 | | pTrcHis Forward | GAGGTATATATTAATGTATCG | 21 | | ITS1 | TCCGTAGGTGAACCTGCGG | 19 | | ITS2 | GCTGCGTTCTTCATCGATGC | 20 | | ITS3 | GCATCGATGAAGAACGCAGC | 20 | | ITS4 | TCCTCCGCTTATTGATATGC | 20 | | pJET1.2F | CGACTCACTATAGGGAGAGCGGC | 23 | | pJET1.2R | AAGAACATCGATTTTCCATGGCAG | 24 | | T7 | AATACGACTCACTATAG | 17 | | T7terminator | GCTAGTTATTGCTCAGCGG | 19 | | T7promoter | TAATACGACTCACTATAGGG | 20 | | T3 | ATTAACCCTCACTAAAG | 17 | | SP6 | ATTTAGGTGACACTATAG | 18 | | M13F-pUC(-40) | GTTTTCCCAGTCACGAC | 17 | | M13R-pUC(-40) | CAGGAAACAGCTATGAC | 17 | | M13F | GTAAAACGACGGCCAGT | 17 | | M13R | GCGGATAACAATTTCACACAGG | 22 | | pGEX5 | GGCAAGCCACGTTTGGTG | 18 | | pGEX3 | GGAGCTGCATGTGTCAGAGG | 20 | | 27F | AGAGTTTGATCMTGGCTCAG | 20 | | 1492R | TACGGYTACCTTGTTACGACTT | 22 | | 518F | CCAGCAGCCGCGGTAATACG | 20 | | 800R | TACCAGGGTATCTAATCC | 18 | | BGH-R | TAGAAGGCACAGTCGAGG | 18 | | CMV-F | CGCAAATGGGCGGTAGGCGTG | 21 | | RVprimer3 | CTAGCAAAATAGGCTGTCCC | 20 | | RVprimer4 | GACGATAGTCATGCCCCGCG | 20 | | GLprimer1 | TGTATCTTATGGTACTGTAACTG | 23 | | GLprimer2 | CTTTATGTTTTTGGCGTCTTCCA | 23 | | pQE-F | CCCGAAAAGTGCCACCTG | 18 | | pQE-R | GTTCTGAGGTCATTACTGG | 19 | | Gal4AD | TACCACTACAATGGATG | 17 | | pBAD-F | ATGCCATAGCATTTTTATCCA | 21 | | pBAD-R | GATTTAATCTGTATCAGG | 18 | | EGFP-CF | AGCACCCAGTCCGCCCTGAGC | 21 | | EGFP-CR | CGTCCATGCCGAGAGTG | 17 | | EGFP-NR | CGTCGCCGTCCAGCTC | 16 |
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What are your preferred methods of purification? |
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We do not have any special preference for purification kit. Conventional column-type purification is good as well as the Exo-SAP purification. |
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How much is it to run the samples with unsatisfactory results again? |
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Retrial for failed ones are provided for free for once.
It is free only when samples and primers left sufficiently and it should be the same DNA and primer match from the first trial.
Note : Free retrial is Not available for Plate sequencing service and 1primer/1plate service. |
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EZ-Seq
