data analysis guide - lnaâ„¢ for microrna research - exiqon
TRANSCRIPT
Data Analysis Guide
For the miRCURY LNA™ Universal RT microRNA Ready-to-Use PCR panels using Exiqon GenEx software Version 3 (February 2014)
Contents
Considerations on Normalization ................................................................................................... 4
Why normalize? ................................................................................................................................ 4
Controls and normalization assays ................................................................................................ 4
Introduction to GenEx ...................................................................................................................... 6
Flow of data analysis ....................................................................................................................... 8
Export the run data from your cycler ............................................................................................. 9
Importing and merging your instrument export files using the Exiqon import wizard .............. 14
Inter-plate calibration ..................................................................................................................... 23
Quality control ................................................................................................................................. 24
Cut off ............................................................................................................................................... 28
Frequency of missing data ............................................................................................................ 29
Call rate plot .................................................................................................................................... 29
Missing data – part 1 ..................................................................................................................... 30
Normalization to global mean ....................................................................................................... 31
Normalization to reference gene(s) ............................................................................................. 32
Missing data – part 2 ..................................................................................................................... 38
microRNAs detected only in a subset of samples ..................................................................... 39
Average RT replicates ................................................................................................................... 39
Converting to relative quantities ................................................................................................... 40
Further data analysis using GenEx ................................................................................................. 42
Using the Data manager ............................................................................................................... 43
Statistical analysis in GenEx ........................................................................................................ 44
Creating a subset of regulated assays ........................................................................................ 46
Dat
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Dat
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a analysis g
name. Ifwhich on As mentautomat #FileNammaking
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Dat
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sample nam
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Dat
20
a analysis g
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Dat
21
a analysis g
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Dat
22
a analysis g
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Dat
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a analysis g
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Dat
24
a analysis g
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ike‐in classifint in the plat
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Dat
25
a analysis g
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Dat
26
a analysis g
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Dat
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a analysis g
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Dat
28
a analysis g
If the sa(a classiidentified
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Dat
29
a analysis g
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Dat
30
a analysis g
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Dat
31
a analysis g
Assays
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