Download AmpFLSTR Yfiler PCR Amplification Kit User`s Manual (PN 4358101C)

Chapter 4 Performing Electrophoresis
Overview
Flowchart
Extract and quantify data
310 Electrophoresis Setup
Set up the instrument
PCR amplify data
Perform electrophoresis
Set up instrument:
- Install and clean pump block
- Install or replace capillary
- Prepare syringes
- Clean electrode
- Recalibrate autosampler
- Fill buffer reservoir
Prepare the samples
• Prepare formamide
size standard cocktail:
- 0.5µL GeneScan® - 500
LIZ® Size Standard®
- 24.5µL Hi-Di™
Formamide per sample
• Vortex
Yes
ABI PRISM®
3100/3100-Avant
ABI PRISM®
310 Analyzer
Matrix file
created?
Create
matrix file
Not sure
Analyze data
4-28
DRAFT
May 25, 2005 9:59 am, 310.fm
No
Display
and check
matrix file
• Into each 0.2-mL or
0.5-mL sample tube add:
- 25µL formamide:
size standard mixture
- 1.5µL allelic ladder
or sample
• Heat @ 95˚C for 3' then
place on ice for 3'
Set up
Data Collection
Software v3.0
If using GeneScan®
Software v3.7.1:
• Preheat the
instrument
• Set up the sample
sheet and injection
list defaults
• Create the
sample sheet and
injection list
Perform electrophoresis
Load samples
Select injection list
If using GeneMapper
ID Software v3.2:
• Preheat the
instrument
• Create the
sample sheet and
injection list
Start run
AmpFlSTR Yfiler PCR Amplification Kit User’s Manual