Create mutagenesis library from existing oligo sets (wizard)

Mutagenesis libraries are collections of DNA oligonucleotides (oligos) that can be used to create variant forms of targeted proteins for a variety of experimental applications (see Learn about mutagenic oligos and mutagenesis libraries). This wizard leads you through the creation of a mutagenesis library based on oligo sets that already exist in your eArray account. You can also create mutagenesis libraries in other ways. See Ways to create mutagenesis libraries.

Before you can use this wizard, you must have one or more mutagenesis oligo sets accessible to you within eArray. To see all of the oligo sets that are available to you, do an oligo set search and leave the search term blank. To create oligo sets, you can use the Create Mutagenesis Oligo Sets wizard, or the Create Mutagenesis Library from Target Sequence wizard.

Overview of wizard

The wizard leads you through several steps, summarized below. For detailed instructions for a specific step, click its name.

To start the wizard

Step 1 – Define Library – In this step, you enter the name of the library and optional descriptive information.

Step 2 – Layout Oligo Sets – In this step, you add or remove oligo sets.

Step 3 – Create Library – In this step, you save the library with a status of Draft, which lets you further edit the library, or Submitted, which submits the library to Agilent Manufacturing, prevents further changes, and lets you request a pice quote for the library. Submitting a library does not place an order for that library with Agilent.

To start the wizard

1. Set the application type to Mutagenesis.
   
   The Home tab of the Mutagenesis application type appears.

2. In the Library Wizards pane on the right side of the screen, select Create Mutagenesis Library from Existing Oligo Sets, then click Next.
   
   The first step of the wizard appears. As you go through the steps of the wizard, click Next to go to the next step. To go back to a previous step of the wizard, click Back (if available). At any time you can click Close to close the wizard. eArray allows you to continue the wizard at a later time.

Step 1 – Define Library

In this step of the wizard, you enter the name of the library and any optional descriptive information.

1. In the Library Name field, type in a name for the library.
   
   The name must be unique for your eArray workgroup. eArray uses the library name to identify the library in search results and other screens.
   

2. If desired, type a description of the library into the Description field.
   
   This description appears when the library is viewed or edited within eArray.
   

3. If desired, type any additional comments into the Comments field.
   
   Your comments are saved with the library and appear when the library is viewed or edited within eArray.

4. Click Next.
   
   The next step of the wizard appears.

Step 2 – Layout Oligo Sets

In this step of the wizard, you add the desired oligo sets to the library.

1. Click Add.
   
   A search window opens.

2. In the Oligo Set Name field on the left side of the window, type in the name, or the start of the name, of the oligo set that you want to add (or leave blank to retrieve all oligo sets in your account).
   
3. Click Search.
   
   The oligo sets found in the search appear on the left side of the window.
   
4. Add one or more of the listed oligo sets to the library.
   
• To add all oligo sets in the list, click Add All.
• To add individual oligo sets, click the sets to select them, then click Add.
• To remove any of the added oligo sets, click the oligo set on the right side of the window (in the Selected Oligo list) to select it. Then click Remove. Clicking Remove All removes all oligo sets.
  
6. Click Done.
   
   The newly added oligo sets are now listed in the Layout Oligo Sets wizard screen.

5. If needed, you can remove an oligo set from the library, or view the oligos in an oligo set.

To remove an oligo set from the library:

1. Mark the check box next the oligo set name that you want to remove.
   
2. Click Remove.
   
   The oligo set is deleted from the library and no longer appears in the Oligo Sets pane.

To view the oligos in an oligo set:

1. Click the oligo set name.
   
   A new window opens displaying details on the oligos and oligo set including oligo sequence
   
2. Click Close to close the window.

6. When you have finished reviewing and editing the library layout, click Next to advance to the next step in the wizard.

Step 3 – Create Library

In this step of the wizard, you save the library with a status of Draft or Submitted.

To save the library as Draft:

1. Select Draft.
   
2. Click Save.
   
   You will be able to make further changes to the library prior to submission.
   
3. Click Close to exit the wizard.

To save the library as Submitted:

1. Select Submitted.
   
2. Click Design Checklist.
   
   A window opens displaying a checklist.
   
3. Read each task on the checklist and confirm that it has been completed by marking the check box.
   
   If you have not completed all tasks, you are not ready to submit the library. Click Cancel to close the checklist window and save the library with a Draft status.
   
4. When you have marked all check boxes, click Done to close the checklist window and return to the wizard screen.
   
5. Click Save.
   
   You have submitted the library to Agilent manufacturing. An Agilent representative will contact you to follow up on your interest. Submitting a library does not place an order for that library with Agilent.
   
   Once you submit a library, you can request a quote for the library as part of the QuikChange Oligo Library Mutagenesis System.
   
6. Click Close to exit the wizard.

See also

Learn about mutagenic oligos and mutagenesis libraries

Overview of working with mutagenesis oligo sets

Ways to create mutagenesis libraries