Agilent eBook - 14
Best Practices with an Agilent Parallel CE System
Sizing under 6,000 bp
throughout a dilution series
×103 A
36 ×103
7
6
5
4
3
2
1
0
32
28
24
20
300 bp fragment
350
300
16
Size (bp)
300
RFU
12
8
LM
UM
4
20
0.06 %
-0.3 %
Table 1. Overview of sizing data for 300 and 1,000 bp fragments over
a concentration range separated on the 5200 Fragment Analyzer
system with the HS NGS Fragment kit (1 to 6000 bp). 1 n = 144.
14
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6,000
700
800
1,000
1,500
2,000
3,000
600
500
400
300
200
100
1,200
1,000
Size (bp)
0.33 %
500
800
600
400
200
0
Size (bp)
6,000
0.41 %
UM
700
800
1,000
1,500
2,000
3,000
3.3
LM
600
1.2
1,000
0
500
992 to 1,001
Range (bp)
Standard deviation
% CV
% Error
1
400
297 to 302
Average (bp)
250
1,000 bp fragment
8
4
500
1,200
300
9971
250
3
200
1,000 bp DNF-474
2991
15.6 31.3 62.5 125
Concentration (pg/µL)
4
100
300 bp DNF-474
7.8
×103
1
Sizing over concentration range
4.5 to 600 pg/µL
3.9
Size (bp)
B
900
RFU
12
150
0
2
16
200
50
×103
28
24
250
100
0
1
The HS NGS Fragment kit has a DNA sizing
range of 100 to 6,000 bp. Fragment sizes of
300 and 1,000 bp were analyzed on the 5200
Fragment Analyzer system with the HS NGS
Fragment kit throughout the concentration
range of the kit (Figure 2A & B). The 300 and
1,000 bp fragments reported consistent
sizing of 299 ± 1.24 bp and 997 ± 3.35 bp
respectively, with a % error below 0.3 %. Both
the 300 and the 1,000 bp fragments had
excellent precision values of 0.41 and 0.33 %
CV respectively, indicating consistent sizing
between the concentration points (Table 1).
0
3.9
7.8
15.6 31.3 62.5 125
Concentration (pg/µL)
Figure 2. Sizing of 300 and 1,000 bp fragments over a concentration range on the 5200 Fragment Analyzer system with the HS NGS fragment kit
(1 to 6,000 bp). A) 300 bp fragment overlay (15.6, 62.5, and 500 pg/µL) and bar graph (3.9 to 500 pg/µL)n = 18. B) 1,000 bp fragment overlay (15.6,
62.5, and 500 pg/µL) and bar graph (3.9 to 500 pg/µL)n = 18. Sizing of fragments under 6,000 bp is consistent, and not affected by concentration
on the 5200 Fragment Analyzer system. LM = lower marker; UM = upper marker.
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Agilent eBook
Table of Contents for the Digital Edition of Agilent eBook
Agilent eBook - 1
Agilent eBook - 2
Agilent eBook - 3
Agilent eBook - 4
Agilent eBook - 5
Agilent eBook - 6
Agilent eBook - 7
Agilent eBook - 8
Agilent eBook - 9
Agilent eBook - 10
Agilent eBook - 11
Agilent eBook - 12
Agilent eBook - 13
Agilent eBook - 14
Agilent eBook - 15
Agilent eBook - 16
Agilent eBook - 17
Agilent eBook - 18
Agilent eBook - 19
Agilent eBook - 20
Agilent eBook - 21
Agilent eBook - 22
Agilent eBook - 23
Agilent eBook - 24
Agilent eBook - 25
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