Biomedical Engineering Reference
In-Depth Information
•
0.5
μ
l of ROX-labeled internal size standard
•
12
μ
l of deionized formamide.
2 Mix by pipetting, incubate at 94
◦
C for 2 min and cool on ice.
3 Start the fragment separation software using the following settings:
•
injection time: 5 s
•
run voltage: 15 kV
•
run time: 30min
•
run temperature: 60
◦
C
•
run voltage: 15 kV
•
filter: C.
ABI-3100 and ABI 3100 Avant:
ss
,
tt
1 Following the PCR reactions, for each reaction mix:
•
0.5
μ
l of ROX-labeled internal size standard
•
8.5
μ
l of deionized formamide
•
1-3
μ
l of the PCR reaction.
2 Pipette this mixture into the injection plate.
3 Seal the injection plate with plate sealing film and incubate at 92
◦
C for 2 min, and hold
at 4
◦
Cfor5min.
4 Start the fragment separation software using the following settings:
run temperature: 60
◦
C
•
capillary fill volume: 184 steps
•
•
pre-run voltage: 15 kV
•
pre-run time: 180 s
•
injection voltage: 3.0 kV
•
injection time: 10-30 s
•
run voltage: 15 kV
data delay time: 1 s
•
run time: 1500 s.
•
ABI-3700:
ss
,
tt
1 Following the PCR reactions, for each reaction mix:
2
μ
l of the PCR reaction
•
0.2
μ
l of ROX-labeled internal size standard
•
10
μ
l of deionized formamide.
•
