Gene Synthesis Ordering Instructions

Ordering synthetic DNA just got a little easier

You no longer have to wait for quotes on standard gene orders. Sigma's new online order form allows you to enter your project details and see instant pricing on constructs from 0.5 - 5kb. Instant pricing means you can place your order right now, and get back to your research knowing your DNA is on it's way.

Every order form you use is a little different, so this page is designed to help you understand the best and fastest way to complete the form.

You may also find the following pages helpful:

Gene Synthesis Quick Ordering Instructions (for those experienced with DNA synthesis orders)

Sigma’s new online order form is designed to allow you to enter your gene synthesis requests very quickly. We intend to provide instant pricing for DNA sequences between 0.5 and 5.0 kb. Once you complete the three sections of the order form, your confirmation page will provide instant pricing, and you can accept these prices by clicking “Add to Cart”, and check-out your cart as with any other online order from Sigma. For sequences of length outside these parameters, or those with unusual complexity, we will provide a message indicating we need to work with GENEWIZ to provide a quote for the request. In order to process this, please add the quote to your cart, and check-out the cart (zero cost) so we can provide the information in your request to GENEWIZ, who will contact you with the quote within 2 business days (usually sooner).

Three sections of the form need to be completed:

  • Construct Information section, where you will provide the sequence name and select a vector.
  • Specifications section, where you will provide the gene sequence (and optional flanking sequences) and restriction enzyme sites to use for the cloning (if applicable – depending on the vector you selected). You will also have an opportunity to indicate your preference for codon optimization.
  • DNA Preparation Information section, where you will indicate if you would like larger DNA preparation sizes. The default is a standard miniprep at no extra charge.

An error will occur if you do not enter information in any of the required fields. All fields displayed to you are required, except the flanking sequences. For more information about how to complete this online order form, see the detailed instructions below.

Gene Synthesis Detailed Ordering Instructions (for those new to ordering DNA synthesis)

Learn about our new order form the easy way – watch our 5 minute video about how to complete the DNA synthesis form.
(insert video here - when it is ready)

Sigma’s online order form for gene synthesis has been designed to make ordering synthetic DNA easy. Access the order form here, and some tips for ordering here. For first-time users, this page describes the use of the online order form in detail. Looking for the shorter version? See the video linked below for a crash course in online ordering. If you have any questions, contact Sigma Technical Support for assistance.

Step 1: Select an Entry Method

Image 1 showing Step 1 (Entry Method).

 

Manual Entry allows you to enter a synthetic DNA sequence order one construct at a time. This is good if you are ordering less than 5-10 constructs. Click on the Manual Entry button to continue (see Image 1).

Multiple Entry allows you to easily enter multiple constructs at once, by either uploading your data using our excel template or by copying and pasting the information in a designated format. Click on the Multiple Entry button to continue.

Step 2: Construct Entry

Manual Entry

The Construct Entry Page for Manual Entry consists of three sections:

Section 1: Construct Information

Choose a name for your construct. If you plan to order more than one DNA construct, give them different names – it helps GENEWIZ during manufacturing and it helps you identify them when you receive your DNA. Use up to 30 letters and numbers – type directly into the field (see Image 2).

 

Image 2 showing Step 2, Section 1 (Manual Entry, Construct Information).

 

There are two options for DNA type – Synthetic DNA Sequence and CRISPR Donor Sequence (see Image 2). Synthetic DNA Sequence is the most common request – these are standard pieces of DNA and this choice allows you the most options in the rest of the form below. The CRISPR Donor Sequence section allows you to order the DNA you plan to use as a CRISPR donor in gene editing projects. Selecting this option does NOT include any CRISPR design – you will need to design the donor sequence first, and enter in exactly the sequence you want. When selecting this option, the form will not allow you to do any codon optimization. If you need help with CRISPR design, consider Sigma’s CRISPR Design Services.

The Expression Vector selection is one of the most important parts of the form – since your choice here will determine how you receive your DNA and how easy it is for you to use when you receive it. You have three vector options free of charge: pUC57Kan, pUC57Amp, and pSF-Core. The pUC vectors are classic vectors that are cloned into by homologous recombination; you can use these expression-ready vectors for expression of your synthetic DNA in E. coli. If you would like to cut your gene out of this vector, you will need to build a restriction enzyme site on both ends of your requested sequence (see Section 2, below). For the easiest cloning into another vector, the pSF-Core is the basic cloning vector that makes up the cornerstone of Sigma’s SnapFast™ Expression Vector Collection.

To save time, you may elect to have your requested DNA delivered in the final expression vector you would like to use in your experiments. For an additional fee, GENEWIZ offers two types of custom cloning, based on your selected vector. The first option is selection of any premium SnapFast™ vector from the vector menu (see image 2) – these represent some of the most popular SnapFast™ Vectors from our Collection. GENEWIZ has them in storage so you don’t need to have them in your lab in order to select them. We have some resources available to help you select the right SnapFast™ Vector for your gene synthesis project – Vector Maps and an article about the SnapFast™ System.

The second custom cloning option allows you to send in to GENEWIZ any vector you would like them to use for your project (see Image 3). In order to select this option, select “Other” for your Vector Selection, provide your vector name, and then answer the following additional questions:

  • Is this vector already at GENEWIZ? Select YES if you have previously used GENEWIZ for gene synthesis and have already sent your vector to them. You will be prompted to supply the previous project ID number – this enables GENEWIZ to find your vector in their archives. Select NO if you have not previously sent your vector to GENEWIZ, and answer the additional questions.
  • Vector Sequence: Enter the sequence of the vector you are sending to GENEWIZ. This allows them to start planning and scheduling the manufacture of your sequence, which requires confirmation of the vector sequence in order to correctly clone your synthesized DNA into the vector.
  • What Antibiotic Selection Marker is used? Choose the antibiotic GENEWIZ should choose for selection of correct clones. This allows them to start planning and scheduling the manufacture of your sequence, which requires confirmation of the antibiotic resistance marker to be used in order to correctly select a clone to use for DNA preparation. If your antibiotic is not on the list to choose from, you can select “other” and write in the name of your antibiotic.

 

Image 3 showing Step 2, Section 1 “Other” vector option (Manual Entry, Construct Information)

Note: your selections in this section of the form determine some of the options available to use for the rest of the form – particularly the restriction enzymes sites to use and codon optimization.

 

Section 2: Specifications

We provide three sections to use for sequence entry (see Image 4). Many customers will use only the Main Sequence box, but some elect to enter their gene sequence into the Main Sequence box, and then use the two Flanking sequences to build in extra DNA sections that aren’t part of the coding region of their gene. For example, DNA spacers or restriction enzyme sites for vectors that usually use homologous recombination (such as pUC57 vectors). For this reason, the Flanking Sequence boxes are marker optional, but the Main Sequence is required. Each sequence entry field has a base counter on the right side.

For all non-pUC57 vectors, you will have the option to select which restriction enzyme site(s) you would like GENEWIZ to use for the insertion of your synthetic DNA. For SnapFast™ Vectors, the options will be limited based on those available in the vector (see vector maps). For other vectors you will be able to choose from a list of over 200 common restriction enzymes and you should choose the enzyme(s) where you want GENEWIZ to cut open your vector to insert the synthetic gene.

 

Image 4 showing part of Step 2, Section 2: Specifications/Sequence Entry (Manual Entry, Construct Information)

 

The sequence preview pane (see Image 5) shows the final sequence your request will generate (not including the vector you chose). We show the sequence in this format: 5’ restriction enzyme sequence + 5’ flanking sequence + main sequence + 3’ flanking sequence + 3’ restriction enzyme sequence. If you want to make any changes to the sequence displayed, you should type in the sequence fields above – not in the preview box.

The sequence preview box is the source of the DNA Properties information we display directly below the sequence preview pane (see Image 5). The base count displayed is the number that your price will be based, and the GC% and AT% calculations enable us to spot potentially complex sequences that we would like to send to GENEWIZ for quote. Even if we determine it is complex, you won’t be charge a complexity fee!

 

Image 5 showing part of Step 2, Section 2: Specifications/Sequence Preview (Manual Entry, Construct Information)

 

You are also given the option to select codon optimization (see Image 6). If you select yes, you will be prompted to provide the target species, and identify if there are any restriction enzyme sites you would like us to avoid. The codon optimization will not be displayed in the sequence preview pane – this work is done by GENEWIZ before they manufacture your DNA sequence.

 

Image 6 showing part of Step 2, Section 2: Specifications/Codon Optimization (Manual Entry, Construct Information)

 

Section 3: DNA Preparation Information

The final section is Step 2 is to identify how much DNA you would like to receive (see Image 7). The standard offer, with no additional charge is the standard miniprep. We also offer midi, maxi, mega, and giga preparation scales, as well as endotoxin-free versions of these larger sizes. Select your preface using the buttons.

 

Image 7 showing part of Step 2, Section 2: Specifications/Codon Optimization (Manual Entry, Construct Information)

 

Notes

Information you enter for the request is automatically saved, even if you navigate away from the page. If you only need one construct, you can click the “Next Step” button to go on to the confirmation page (see Image 7). Alternatively, you can use the button for “Add A Construct” to enter the details of your next construct/sequence request. When completed, use the “Next Step” button to move to the confirmation Step.

If you would like to view all the constructs on the order request, and/or delete one or more constructs, use the “View All Constructs” button. If any of your constructs contain errors, you will be prompted to correct those before adding the order to your cart (see Image 8).

 

Image 8 shows the “View All Constructs” pop-up.

 

Multiple Entry

The multiple entry of gene synthesis requests is available in two ways: file upload or copy & paste (see Image 9).

 

Image 9 shows the screen with two multiple entry options: file upload and copy & paste.

 

In order to use the file upload feature, first download the Bulk Gene Template and complete the information in the required fields. Notice the instructions and tips provided across the top row of the template. Save the completed form to your computer, and use the “Select File” button to choose the saved file and the “Submit” begin the upload process. Once uploaded, the construct information will be input into the online form and you can proceed to the “Next Step” – the Confirmation Step (last page). Any errors from the upload will be displayed to you can correct them manually before using the “Add To Cart” button to proceed with check-out for order completion.

In order to use the Copy & Paste bulk entry function, prepare your request data in the format shown in the order form:

Construct Name,DNA Type,Expression Vector,Vector Name,Is this vector already at GENEWIZ?,Vector ID,Vector sequence if vector in not at GENEWIZ,Antibiotic Name,Antibiotic Name if Other selected from the list,5' Flanking Sequence,Main Sequence,3' Flanking Sequence,5' Restriction Enzyme,3' Restriction Enzyme,Codon Optimization,Expression System,Restriction Sites,DNA Preparation Size,DNA Preparation Type

Once complete, click the “Submit” begin the upload process. Once uploaded, the construct information will be input into the online form and you can proceed to the “Next Step” – the Confirmation Step (last page). Any errors from the upload will be displayed to you can correct them manually before using the “Add To Cart” button to proceed with check-out for order completion.

Step 3: Confirm Order

Once you navigate to the Order Confirmation section of the online order form, you can review the details of all your constructs together, and see the price of each construct request individually (see Image 10). You can delete any constructs you do not want to order, or click the link to the construct to change the details. If there are no errors noted, you are able to “Add to Cart” and check-out your cart, as with any other Sigma online order. Once you check-out, we start the process of sending the request to GENEWIZ, where they will confirm the order is complete and start the planning and scheduling of the gene synthesis request.

 

Image 10 showing part of Step 3 (Order Confirmation)

 

Step 4: Check Out Your Order (or Quote)

TBD


Looking to learn more about gene synthesis? Click here to go back to the basics before you place your order.

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