Agilent eBook - 13
LM
6,000
3,000
1,500
2,000
1,000
700
800
UM
600
500
400
300
200
1
Average smear size 312 bp
100
RFU
Best Practices with an Agilent Parallel CE System
5,000
4,500
4,000
3,500
3,000
2,500
2,000
1,500
1,000
500
0
Size (bp)
B 6,000
UM
LM
5,000
LM
Peak size
350 bp
RFU
4,000
UM
3,000
2,000
Peak size 300 bp
1,000
Size (bp)
6,000
3,000
1,500
2,000
1,000
700
800
600
UM
LM
5,000
LM
RFU
4,000
Average smear size 312 bp
UM
3,000
Average smear size 488 bp
2,000
1,000
6,000
3,000
1,500
2,000
1,000
700
800
600
500
400
300
200
100
1
6,000
3,000
1,500
2,000
1,000
700
800
600
500
400
300
200
100
0
Size (bp)
B
500
Size (bp)
6,000
1
5,500
5,000
4,500
4,000
3,500
3,000
2,500
2,000
1,500
1,000
500
0
400
300
200
100
1
6,000
3,000
1,500
2,000
1,000
700
800
600
500
400
300
200
0
100
5,000
4,500
4,000
3,500
3,000
2,500
2,000
1,500
1,000
500
0
1
RFU
A 5,500
RFU
DNA smears were separated on the 5200
Fragment Analyzer system with the HS NGS
Fragment kit in order to compare peak and
smear sizing. DNA smears or libraries with
a typical bell curve or a uniform distribution
on both sides of the highest peak of the
sample, reported similar peak and smear
sizes (Figure 1A). In contrast, DNA smears
with a larger distribution after the highest
peak reported a larger smear size compared
to the peak size, as expected (Figure 1B).
In the same respect, DNA smears with a
larger distribution before the highest peak will
report a smaller smear size compared to the
peak size. Therefore, the most accurate sizing
for a smear or library sample can be achieved
with the Smear Analysis tab on ProSize.
Size (bp)
6,000
RFU
UM
Figure 1. PeakLMand smear size of DNA libraries analyzed on the 5200 Fragment
Analyzer system with the HS NGS Fragment kit (1 to 6000 bp). (A) A DNA library
5,000
with
a bell‑shaped curve will report similar
peak
and
smear
sizes.
(B)
A
DNA library with a larger distribution of fragments to the right of the highest peak will
Peak size
have
a larger smear size than peak size.
analysis is recommended for sizing of all DNA smears and library samples.
350 Smear
bp
4,000
3,000
2,000
1,000
6,000
3,000
1,500
2,000
1,000
700
800
600
500
400
300
200
100
1
0
Size (bp)
6,000
UM
LM
5,000
13
| GENengnews.com
3,000
Average smear size 488 bp
2,000
1,000
0
1
RFU
4,000
http://www.GENengnews.com
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
https://www.nxtbookmedia.com