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A convenient system for highly specific and sensitive detection of miRNA expression
Xiangqi Li, Minjie Ni, Chaobao Zhang, Wubin Ma, Yonglian Zhang
SUPPLEMENTAL DATA
A
B
C
miR
-138
miR
-99b*
miR
-125
a-3p
miR
-291
miR
-292
-5p
miR
-295
miR
-326
miR
-308
5
NO miR
-122
Mar
ker
miR
-138
miR
-99b*
miR
-125
a-3p
miR
-291
miR
-292
-5p
miR
-295
miR
-326
miR
-308
5
NO miR
-122
Mar
ker
miR
-138
miR
-99b*
miR
-125
a-3p
miR
-291
miR
-292
-5p
miR
-295
miR
-326
miR
-308
5
NO miR
-122
Mar
ker
Fig. S1 Detection of miRNAs under different conditions by stem-loop PCR.
50bp100bp
50bp100bp
50bp100bp
Fig. S2 The sensitivity of the detection of miR-7578 and distinguishing miR-7578 from its precursor by northern blotting.
Rno-miR-7578
Pre-miR-7578
10 fmol
1 fmol
1 amol
1 zmol
JEG
-3 C
ell
MG
-63
Cell
U-2 O
S cel
l56
37 C
ell
T24 C
ell
Rno-miR-29a by aLHCD
Rno-miR-29a by northern blotting A
B
Fig. S3 Detection of miR-29a in different cell lines by different methods.
Fig. S4 Quantitative detection of miRNA levels using U6 as an internal control by LH-PCR.
Initial relative quantity
I1 I2 I8
Re
lativ
e q
ua
ntity
by
LH-P
CR
0
2
4
6
8
10
Fig. S5 Evaluation of LH-PCR amplification efficiencies of miRNA and its precursor.
RNA
Pre-miR-122 miR-122Re
lativ
e a
mp
lific
atio
n e
ffe
cie
ncie
s
0
20
40
60
80
100
120
miR-200b family
miR-200a miR-200c miR-200bRe
lativ
e a
mp
lific
atio
n e
ffe
cie
ncie
s
0
20
40
60
80
100
120
Fig. S6 Evaluation of LH-PCR amplification efficiencies of miRNA and its family members.
miR-29a
miR
-29a
miR
-29b
miR
-29c
Tissue RNA
Fig. S7 Detection of miR-29 family members by conventional Northern blotting.
Rno-miR-138
pre Hela liver kidey lung brain
Fig. S8 Detection of miR-138 and its precursor from tissue samples by stem loop-PCR .