Start SEQUENCHER

Drag sequences to SEQUENCHER window

Open files by double clicking

Only use files with good signal

Find restriction site of vector (eg. EcoRI GAATTC) with FIND option

Crop vector sequence

Open chromatogram

Find where good sequence ends or poly A tail starts

Crop rest of sequence

Export sequence as Pearson text

One file:

• copy sequence into window at NCBI webpage

• do basic blastn nr search

Multiple files:

• start SEARCH LAUNCHER by dropping sequence files onto the camel symbol

• type DNA

• type 10 (BLASTN NR) or 12 (dbEST)

• results are returned into the SEARCH LAUNCHER directory

The mapping macro generates one map per freezer box, but it’s quickest to pull the data out by one freezer at a time.

• Log in to Freezerworks. If you don’t know how, you should get help from Tara (queen of the Freezer people)
• From the Samples menu, choose Search by Location
• Select the Freezer you want to export. Leave the Rack, Box, Position boxes blank to select all of the samples in the freezer.
• Open the Import/Export menu and choose Export Data.
• Double-click the Map Maker export format.
• Make sure the Field Delimiter is still tab and the Record Delimiter is return and that Include Header Record is checked.
• Click the Save Export button and note where you save the file for the next step

Creating the Freezer Data File in Excel

Open a new Excel worksheet

Choose Data Get External Data Import Text File

select the freezerworks file you created in step 1

click Finish to accept the defaults and import the data

Fill the empty Subdivision 5 column with spaces:

• enter a space in the top of the column (probably cell C2)
• select that cell and everything below it (shift + cmd + down arrow)
• choose Edit Fill Down

Sort the data by position

• Select all of the data (cmd + A or edit select all)
• Choose Data Sort
• Sort by Subdivision 1 Position, then Subdivision 2 position, then Subdivision 3 Position with a Header row

Create a Map of the Data for a Box

• If you plan to save, create a copy of the MapMaker Blank.xls file. You can find it in the ‘Freezerworks Webinars’ folder on transfer if necessary.
• Copy and paste the rows corresponding to a single box from the freezer worksheet into the ‘Paste Data’ worksheet of the Map Maker file. The shelf column should be blank because we don’t use that subdivision.
• Press ctrl+a to activate the magic. The data in the Paste Data tab will be transformed into a map of the box on the Map tab.
• Print or save as you like. (you may need to redefine the print area to include the line numbers)