supplemental figure 1 no substrate - university of...
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2‘,3’ cyclic phosphate substrate no sub
strate
1 2 3 4 5
Supplemental figure 1
Reverse transcription oligonucleotide (adaptor complementarity)
PCR oligonucleotide (transcript complementarity)
0 20 40 60 80
005
001051
002052 Ala
AGC (n=353)CGC (n=85)TGC (n=64)
0 20 40 60 80
0004
0080021
ArgACG (n=1403)CCG (n=632)CCT (n=170)TCG (n=503)TCT (n=44)
0 20 40 60 80
0001
002003
004
AsnGTT (n=528)
0 20 40 60 80
00001
000200 03
00 04 AspGTC (n=4855)
0 20 40 60 80
001
0203
0405 Cys
ACA (n=1)GCA (n=144)
0 20 40 60 80
002
0406
08001
GlnCTG (n=221)TTG (n=279)
0 20 40 60 80
0005
00010002
GluCTC (n=6794)TTC (n=5919)
0 20 40 60 80
0001
003005
GlyCCC (n=2512)GCC (n=1218)TCC (n=427)
0 20 40 60 80
0002
004006
HisGTG (n=2050)
0 20 40 60 80
002
0406
0800 1 Ile
AAT (n=111)TAT (n=0)
0 20 40 60 80
005
001002
LeuAAG (n=68)CAA (n=126)CAG (n=571)TAA (n=41)TAG (n=29)
0 20 40 60 80
0002
0060001
LysCTT (n=2346)TTT (n=946)
0 20 40 60 80
005
001051
MetCAT (n=720)
0 20 40 60 80
002
0406
08021 Phe
GAA (n=268)
0 20 40 60 80
005
001051
ProAGG (n=31)CGG (n=211)TGG (n=361)
0 20 40 60 80
005
051052
0 53 SeC(e)
tRNA position
TCA (n=635)
0 20 40 60 80
02
46
801 Ser
AGA (n=5)CGA (n=7)GCT (n=33)TGA (n=5)
0 20 40 60 80
005
001051
00 2
Thr
tRNA position
AGT (n=195)CGT (n=348)TGT (n=33)
0 20 40 60 80
002
0406
0800 1 Trp
tRNA position
CCA (n=197)
0 20 40 60 80
001
0203
04 TyrGTA (n=45)
0 20 40 60 80
0002
004006
00 8 ValAAC (n=424)CAC (n=1764)TAC (n=598)
sdaer gnicn euqeS
tRNA position tRNA position
Supplemental figure 2
sdaer gn icneuqeS
sdaer gnic ne uqeS
sd aer g nic neu qeS
sdaer gnicne uqeS
1. TPT1 + NAD2. RNA fragmentation
+
p5’-
5’-p
5’-App
p5’- p
5’- p
ddC- 3’
ddC- 3’
ddC- 3’
ddC- 3’
+
+ ATP
PO4
+ Appr>p
1. tRNA Ligase2. Gel purification
Reverse Transcriptase
5’- p ddC- 3’- 5’3’-
RNA hydrolysis
5’- - 3’ 5’-App ddC- 3’+
5’- ddC- 3’
1. T4 RNA Ligase2. Gel purification
PCR and High-throughput Sequencing
Supplemental figure 3