FREQUENTLY ASKED QUESTIONS

Getting Started
An online account may be set up by clicking on the ‘Client Login’ tab on our home page, or by clicking here.

Once you are at the Client Login page, click ‘Register Here.’ You will be asked to submit basic information such as your name and institution, address, phone number and email address. You will also be asked to enter a Username and Password. This is how you will login to your account in the future.

Once you have entered the required information, an account will be opened and you will be able to place new orders.

Yes, we do offer a free trial that includes three free sequencing reactions and free shipping. On our home page, click Free Sequencing Trial at the bottom of the page. You will be asked to enter your sample information, including the names of both your templates and primers, as well as the type of DNA you wished to have sequenced.

Once you have entered your sample information, you will be directed to open a new account with us. You will be asked to submit basic information such as your name and institution, address, phone number and email address. You will also be asked to enter a Username and Password. This is how you will login to your account in the future in order to place new orders as well as access your data.

If you are at an institution in which there is an ACGT, Inc. dropbox, please drop off your samples in the box before the scheduled pick-up time. The samples will be collected by our courier and shipped to our facility. If your institution does not have a dropbox, we will send you a shipping label so that you may send your samples to us free of charge.

Please submit a quote request on our website or contact us at 1-800-557-2248 to obtain pricing information.
You would have to register with us by creating an account online in order to place an order.

Once you have created an account, log in to the account and click on ‘New Orders.’ From there, click on the appropriate service in order to place a new sequencing order.

We retain all the account records on file. However, if you have not used us since 2006, you are required to open a new online account prior to placing an order. For others, login to your account as before. Once you login, click on ‘New Orders.’ From there, you will be able to place an order for the service of your choice.
Choosing A Service
The Premium Service does not require you to quantitate your template or primer DNA. Once we receive your template and primer DNA, we will determine their concentrations via agarose gel and/or Nanodrop and optimize them for the sequencing reactions.

The Standard Service allows you to submit your template and primer DNA in separate tubes. Under this service, we ask that you provide the concentrations of both the template and primer DNA, which we will then use in order to optimize the sequencing reactions.

The Low Cost Option requires you to premix your template and primer in one single tube according to our sample submission requirements. If you are using one of our free Universal Primers, we will add it for you at no additional charge. Individual banked primers cannot be used for this service.

If you need help selecting a service, feel free to call us at 1-800-557-2248.

A Single Pass Primer Extension order is one in which we simply sequence your samples with the designated primers and supply you with individual sequences.

Full Sequencing Analysis is usually performed when the insert size is greater than 2 kb. Primers for this type of sequencing may either be provided by you, or we can design and synthesize the primers for you. Typically, a consensus sequence, an alignment file and a report comprising of the results and variant report would be sent to you. Please contact us for a quote before placing an order. If you are not interested in getting details reports, feel free to use our Single Pass Primer Extension service.

If you need help selecting a service, feel free to call us at 1-800-557-2248.

We guarantee at least 750 base pair per sequencing reaction, though we are usually able to supply sequencing reads of 1 kb or greater.
Which service you choose depends on what exactly you need. If you need to be provided with individual sequences that you will align yourself, you may use our Single Pass Primer Extension service. We also provide an option of providing you with consensus sequence using our Single Pass Primer extension service at an extra charge. If you are interested in this, make sure this option is selected on the online order form.

If, however, you want details reports on sample processing, data analysis and variant report, please select our Full Sequencing Analysis service.

If you need help selecting a service, feel free to call us at 1-800-557-2248.

You could choose either our Single Pass Primer Extension service or Full Sequence Analysis based on the deliverables to would want. Please refer to the question above for more details. If you need help selecting a service, feel free to call us at 1-800-557-2248.
Yes, we are able to sequence such samples using one of our modified protocols. Be sure to select either High GC or Hairpin when placing an online order.

Please note that additional charges apply for these protocols. Please submit a quote request on our website or contact us at 1-800-557-2248 to obtain pricing information.

Direct Colony Sequencing Service
We do not recommend suspending the colonies/culture in water as this can cause unfavorable sequencing results. We suggest using PBS pH 7.2 to 7.5, or TES (TrisHCl 10mM, EDTA 1mM, NaCl 150mM) pH 7.2 to 7.5 for sample suspension.
The turn around time for this service is 2 days from sample receipt.
High-Throughput Sequencing Service
In order to qualify for our High-Throughput service, 48 or more sequencing reactions must be submitted in a single order. Samples need to be submitted in either 8-strip PCR tubes or a 96 well plate. Also, the High-Throughput online order form will need to be filled out. The discounted rate will not be applied if a Single Pass Primer Extension order form is used.
Sample Submission
Yes, you can ship your samples for delivery on Saturday. ACGT, Inc. is open on Saturdays for sequencing and the data will be uploaded for you on Monday.
We recommend naming your samples in a simple, unique way. Long names can often be difficult to write on a tube and abbreviations of these names can lead to human error. Please make sure that the name on the tube completely matches that on the order form.

It is also important to keep in mind that our software used to analyze the samples only accepts letters, numbers, underscores and dashes in the sample names. Please try to avoid using any other symbols in the name of your samples.

The names of the samples should be written on the top of the tube in black marker. Please avoid using other colors as these are often difficult to read.

We strongly recommend sealing your plates with strip-caps. While sealing with plate seals or foil may appear to be sufficient, we often find that leakage occurs during shipment that can lead to contamination. We also recommend wrapping the plate in parafilm to ensure that the strip-caps stay in place during shipment.
Samples may be submitted in either 1.5 mL tubes or strip tubes. If you are submitting your samples in strip tubes, please make sure that they are clearly labeled.
While the sample may sequence well when suspended in TE buffer, we strongly recommend suspending the sample in water. Buffers often contain salts and other contaminants that may interfere with the sequencing and lead to poor sequencing results.
We strongly recommend determing the concentration of DNA by running it on an agarose gel and comparing it to a standard of known concentration. While the concentration may also be determined by using a Nanodrop, the agarose gel method is more accurate.
We highly recommend purifying your DNA before submitting it for sequencing. Unpurified DNA often contain salts and other contaminants that may adversely affect the sequencing reads.

Column purification is sufficient for plasmids and PCR products with one single robust band. For PCR products that contain multiple bands, gel purification of the band with the desired size is required. We do recommend following gel purification with a round of column purification. Please note we recommend eluting the samples in nuclease free water instead of the elution buffer provided with the kit. Please refer to our Sample Submission Requirements for more information

We can also perform column or gel purification of your sample for a nominal charge.

Samples and primers are normally stored for two weeks after they have been received.
We do offer a free primer banking service that allows you to store your primers and/or samples at our facility longterm. Please indicate on your order form if you would like to take advantage of this service.
The amount of DNA needed varies depending on the type of DNA you are sending. Please refer to our Sample Submission Requirements for more information.
A notification email will be sent to you when your samples have been received.
Yes, we can have primers synthesized for you. Simply enter the primer name and sequence in the order form and the primer will be synthesized. Please note that in order for primers to be available the day we receive your samples, orders must be placed by 1 pm CST.
DNA Sequencing Results
When you download the data, you will receive two files – a .seq file and a .ab1 file. The .seq file is a text file that can be opened using a word processor such as Notepad or Microsoft Word.

The .ab1 file is a chromatogram and may be opened using a variety of programs. If you do not currently have a chromatogram viewer, we offer several for download. Please click here for more information.

The data can be interpreted by viewing both the chromatogram as well as the sequence file. We always recommend looking at both files in order to view any discrepancies between the two.
When you receive the data is determined by which service option you have chosen. If you selected our Same Day Service, you will be provided with data by 6 p.m. CST the day we receive your samples. If you have selected our Standard Service, you will be provided with data by 12 p.m CST the day after we receive your samples.

The data will be uploaded to your personal account. To retrieve the data, you will need to login to your account. Once you have logged in, click on ‘My Account’ followed by ‘My Order.’ From there, you will be able to download the data for the desired order number.

Data remains uploaded to your account indefinitely. Therefore, you can easily access data from previous orders by vising ‘My Orders.’
Data will be available through your online account indefinitely.
This most often happens when the data you received from your sample is unusable. When such data is generated, the analyzer often generates a “junk” sequence that is not at all reliable. When such sequences are blasted, they may align with genes other than the one you are interested in.
While PCR primers may occassionally work for sequencing, this is often not the case, for a couple reasons. First, the specifications for PCR and sequencing primer design can vary. Therefore, if a primer was designed according to PCR specifications, it may not work for sequencing.

Second, the PCR products generated may have lost the “primed” region. In other words, the region of DNA that the primer binds to may not be present in the PCR product.  In order to overcome this possibility, we advise using nested primers or designing new internal sequencing primers.

Miscellaneous
You do not need an official quote to place an order. All we need is the name and contact information of your purchasing agent.
Yes, we do offer a Primer and Sample Banking service that allows you to store primers and/or samples longterm. Please be sure to mention on the order form that you would like to take advantage of this service.
Samples and primers are normally stored for two weeks after they have been received.
We do offer a primer banking service that allows you to store your primers and/or samples at our facility longterm. Please indicate on your order form if you would like to take advantage of this service.