Guidelines for Sample Submission to Sequencing Core Facility
Please contact the Sequencing Facility Manager when sending samples. There are three options for sending samples:
- Schedule an appointment to bring samples to the facility. Email the facility the lab manager, Ryan Percifield at (304) 293-4542 or (304) 293-8051.
- Send samples by Campus Mail to Post Box 6057. In this case samples should be packed properly for shipping and the facility manager should be notified in advance.
- Get the lab manager to pick them up from you. This requires that you be nice to Ryan, which is really pretty easy.
Ordering Sequencing or Genotyping Service
This Genomics Core Facility website allows you to upload a file (comma or TAB-delimited text file) that specifies sequencing information. Additionally, you can also upload a Gel image. Below you’ll find information about proper formatting and example files. Ordering and sample information uploads can be accomplished on the Sequencing Orders page
Proper File Format
The purpose of the flat-files is to specify well information for your sample sequencing. The genetic sequencer uses a plate that contains 96-wells. Of those 96 available wells, your flat-file must specify which wells to use, their contents and parameters. You can specify up to 96-wells per work order and thus per flat-file. You cannot use multiple flat-files for one order. If you need more than 96 wells, please submit multiple orders to do so.
Working with Wells
Each well on the plate has a specific alpha-numeric value associated with it. There are eight alpha-columns (A-H) and twelve numeric-rows (1-12) per plate for a total of 96 wells (A01 through H12). The specification is alpha-column first and numeric-row second, like A05. The system allows you to specify which well to begin on but MUST be an ODD numeric-row and must be an A like A11.In order to start on a specific well, you must upload a flat-file. You cannot manually enter samples and start on arbitrary well. In all other cases, the system assumes you’re starting on well A01.
You can optionally omit the Well information from the flat-file. The system then assumes that you are starting on well A01. You can additionally specify only the initial starting well and omit the others, the system will increment the remaining wells for you. If you specify an arbitrary starting well, you must ensure you stay within the 96-total wells of the plate, otherwise the flat-file will be rejected. Again, if you wish to specify arbitrary starting well it must be one of the possible Axx wells, xx being an odd number.
The system expects the sequencing well information in lines and ignores blank lines. It also expects well-specific information in columns as outlined below. The columns can be separated by commas or by TABs.
Here are the default required columns:
- Template Name
- Template Concentration (ng/uL)
- Name of Primer
- Primer Concentration (pmole/uL)
- Vector Name
- Size of Insert (bp)
The column names above in general should be specified in the flat-file upload but could be entirely omitted if you choose. If you include the column names, you must make it the first non-blank line of the file and you must spell at least the first two columns exactly as shown above (excluding the units in brackets) in order for the system to properly recognize them. That is, you must specify at least the first two columns for it to recognize the line as the column header. Therefore, if you name your actual well data like the column header on the first non-blank line, the system will treat it as the column header. Moreover, whether you specify the columns or not, the order in which the columns appear above, from left-to-right, are the order the system expects for your data. That is to say, you cannot rearrange the column header and expect the system to accept it.
Note: The system ignores blank lines but DOES NOT ignore extra TABs in-between columns. That is, if there are multiple TABs between column names or data, it will count each TAB and possibly cause the upload to fail.
As mentioned, the system is rather dynamic in that you can omit various pieces of information and the upload will still work. For example, you can sometimes omit the Wells column if you wish, the system will assume you are starting with well A01.
You can specify only the Wells and Template Name when running Option C from the Order and Services page. If you run Option A or B, however, you must specify the all the column information in the flat-file, even if you upload a Gel image (with the exception of omitting alpha-numeric Well data). If you specify a Gel image and run Option A or B, you can fill-in fake data as placeholders. We recommend using the examples below as your starting point since they already contain fake/dummy data.
When specifying the Template Name in any case, you MUST specifty unique names for each well. The only exception is when specifying a blank well as described below (that is, you can have multiple Template Names with “blank” specified).
You can specify blank wells (in-between actual, used wells) if you wish. In order to do so, specify Blank or F for the Template Name in the file. Also note that the specification is not case-sensitive, so blank or f would work also. If you wish to specify blank wells, you must upload a flat-file. Blank wells before or after the used wells in the plate are taken care of for you.
Example Flat Files
Any of the following samples will work with Option C (both Fragment and Non-Fragment Analysis) in the Orders and Services section. However, only the examples that contain all columns (Temp. Name, Temp. Conc., Name of Primer, etc ) will work with Options A and B.