Bio-Rad Fluorescent Assay Kits - Manual (Page 4)

2.2 Using the CE dsDNA 100 Base Pair
Ladder
The CE-LIF dsDNA 100 bp ladder is a mixture of 10 double-
stranded sequences ranging in length from 100 bp to 1,000 bp in exact
100 bp increments. Under the conditions described below, the standard
will exhibit 10 major peaks (see Figure 1). Pipette 20–50 µl of the
CE-LIF dsDNA 100 bp ladder into a 500 µl sample vial. Routinely, the
CE-LIF 100 bp ladder should be stored at 4 °C. However, longer term
storage at -20 °C may increase shelf life.
Fig. 1. Separation of 100 bp ladder.
3
Section 2
CE dsDNA 1000 Fluorescent Detection
and Kit Components
The kit contains all the reagents for 80 setups, with 30 runs per
setup using the BioFocus LIF 2 detection system.
• Two BioCAP DNA analysis capillaries, 50 cm x 75 µm ID x
375 µm OD
• CE dsDNA run buffer in 2x TBE, 60 ml (2 bottles)
• CE-LIF dsDNA 100 bp ladder, 200 µ l at 1.0 µ g/ml in TE
buffer, pH 8.0
• CE Grade SYBR Green I, 200 µl (Since this fluorescent dye
is light sensitive, do not leave exposed to light when not in
use. Store the fluorescent dye at 4 °C or less.)
• Instruction manual
2.1 Capillary Preparation and Use
The BioCAP DNA analysis capillary supplied with this kit is
designed for use with the BioFocus LIF user assembled capillary
cartridge. For information on capillary installation, refer to the
instructions included with the BioFocus LIF cartridge assembly kit,
catalog number 148-3054. For use with this kit, the capillary car-
tridge can be assembled with 24 to 44 cm (total length) of capil-
lary installed.
2
Minutes
RFU
100 bp
500 bp
700 bp
800 bp 900 bp 1,000 bp
200 bp
300 bp
400 bp
600 bp
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0.9
0.8
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0.6
0.5
0.4
0.3
0.2
0.1
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-0.1
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