Mutation Explorer™

Quick Users Guide

1. Installation: Click the setup file of Mutation Explorer, the software will be installed to C:\program files\mutation Explorer\ unless you specify the differently.

2. Open sample data at the directory of C:\program files\mutation Explorer\hetero….

SoftGenetics Inc.State College, PA 16803Phone: 814-234-8483

Email: MutationExplorer@att.netwww.softgenetics.com

Open Gene Information file

1. Click this fold will lead you to open file dialog box.

2. Click here will ask you to go to the directory to open the text file. You may go to c:\program files\mutation Explorer\hetero\gene_ infor.txt or select your own data for importation

3. Click “Add” will allow to input Genbank file *.seq by inputting c:\program files\mutation Explorer\hetero\*.seq. Input ab1 or scf sequence trace files to reference and sample areas of dialogue box in a similar fashion.

Run the program4a. ME flow-chart (data input, contig, mutation calculation, report, and restart). You may click one by one or click report to get final results.

4b. Click “mutation report output” and analysis is completely automated finishing with Mutation

Report in seconds

Mutation Report

5. Mutation Report: Red color, lower score; blue color, high score greater surety of mutation call.

6. Click any text of the report will lead you to the

electropherogram, to confirm

mutation call

Mutation Explorer Layout

File manage-

ment

7. Sequence texts of ref, sample, genbank.

* means inconsistency

8. Amino acid sequence from genbank

Reference electropherogram

Sample electropherogram

9. Mutation Electropherogram. Mutation if it is a peak. Indel if

center line turns red.

Genbank3’5’

Mutation peak table

Hot Keys

Draw a box from left to right with left mouse to zoom-in an electropherogram

Draw a box from right to left to zoom-out.

Hold left mouse and drag to move across

electropherogram

Click the mutation peak table to zoom-into the mutation peak

Hold Ctrl and click left mouse to manually select genbank, reference, or a mutation peak

Trace quality

boundary

Mutation Parameters

Intensity: mutation peak height. Area: the area of the mutation peak (x1000).

Dropping factor: The percentage of intensity drop

from reference to sample with respect to neighbor bases. Overlapping factor: the mutation peak relative to

reference and sample peaks.

Sn-ratio: signal-to-noise ratio. Mutation Score: accuracy score in logarithm scale.

Comments: user may add comments.

ME parameter settings

Click the icon to change the red peak thresholds