Sequence Detector

SEQUENCE DETECTOR (B407) - Brown Lab

7900HT Fast Real-Time Sequence Detector (B407)

Rules and Guidelines

BEFORE USE:

1. New user MUST be trained by the captain or present users

2. Log user information. To sign up to use this instrument,

a) Go to the calendar at www.yahoo.com.

b) The user id is “sequencedetector”.

c) The password is “patbrown”.

3. Have your plate ready prior to time of instrument operation.

USER INSTRUCTIONS:

1. Depress start button located at the lower left corner of the face of the instrument.

2. Select Brown Lab icon and then on the desktop, select SDS2.3 shortcut. A login window will appear with “administrator” as the password; select “ok” because there is no password.

In the File dropdown list, select new plate, and then select the appropriate plate format for your experiment

a) Absolute Quantification (AQ) used with standard curves

b) Relative Quantification (RQ)

c) Allelic Discrimination (ΔΔCt)

3. Fill out the plate format:

In the “Setup” tab:

a) Select your detectors by selecting the button “Add Detector” at the bottom/center.

b) Assign the detectors to their appropriate wells and select their function Unknown/Standard/NTC

c) You can name the wells later. You can not copy from one plate and paste to another. Instrument will not run using a copy.

e) Select “Instrument, Thermal Cycler” subtab:

f) Check that the volume shown in the upper right corner is appropriate.

g) Check that the cycle, temperature and time presets are appropriate

h) Select “Instrument, Real Time” subtab:

i) Select the button “Connect to Instrument”

f) Wait until the “Instrument Status” window says Connected and highlighted in green.

4. Select the Open/Close button

a) The door opens and the arm rotates out to receive your plate.

b) Place your plate such that well A1 is in the upper left corner.

c) Select open/close button and the door will close.

5. Select Start Run button.

TROUBLESHOOTING

Absolutely do not touch door during the run or solenoid inside the door on the left. Your run will fail.

If there is a very large clanking sound at onset of the run, photo-optic head is not aligned to your plate so you should notify captain asap.

If instrument shows one second or one repetition left in the run and has been running for minutes or hours, save run, close program, restart program, open your file, analyze and your data will boot up.

MANUFACTURER

Applied Biosystems

FULL MANUAL: File:ABI Prism 7900HT User Guide.pdf

FOR SERVICE

Service engineer is Janette Panknin

For tech support or to open a service call, 1-800-762-4001