Pet

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Shahrzad Mirbaha

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GIS-PET
GIS full-length cDNA
(pGIS4)
biotinylated CapTrapper

GIS-PET



(flc-DNA)



cDNA mRNA
diol
5Me-dcTP

cDNA

GSU1

A









cDNA
cDNA

,TCCGAC
PET

cDNA
NOT1 

Mme1 
T4 



3
CDNA

Cap – Trapper

diol
mRNA cap
5Me-dcTP




STREPTAVIDIN
A

GSU1
CTGGAG

STEP1

AAAAAAAA
CTGGAG

STEP2

STEP3
T16

GCGGCCGC :GIS N5 GCGGCCGC:GIS N6

STEP5

NOT1
NOT1

M/B/S
STEP6
PGIS4
GIS-full length cDNAlibrary

TCCGAC

STEP7

Mme1

Mme1 digestion and end polish using T4 polymerase

AA
XX

NN

TT

20bp
Self-ligate , propagate in bacteria
Extract single- PET plasmid DNA
STEP8

18bp
GIS single PET library

AA

M/B/S

STEP9

release and purifry 50-bp PET
b

M
b

Concanate PETs into vectore
STEP10

GIS PET library

STEP11

Sequencing and mapping
GIS PETs to genome

Gen1

Gen2

Gen3

PET
transcriptome
Genome annotation





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Pet

1. PET(Paired End di-tags)

4. PET cDNA cDNA 3 , 5  18bp  PET 

5. PET  GIS – PET CHIP – PET MS-PET 

6. GIS-PET fl-cDNA  GIS-PET 

7. GIS-PET GIS full-length cDNA (pGIS4) biotinylated CapTrapper GIS-PET  (flc-DNA) 

8. GIS-PET PET PET mRNA Cap Trapper 

10.

11. cDNA mRNA diol 5Me-dcTP cDNA GSU1 A     

12.  cDNA cDNA ,TCCGAC PET cDNA NOT1   Mme1  T4   

13. 3 CDNA Cap – Trapper  diol mRNA cap 5Me-dcTP   STREPTAVIDIN A GSU1 CTGGAG

14. STEP1 AAAAAAAA CTGGAG STEP2 STEP3 T16 GCGGCCGC :GIS N5 GCGGCCGC:GIS N6

15. STEP5 NOT1 NOT1 M/B/S STEP6 PGIS4 GIS-full length cDNAlibrary

16. TCCGAC STEP7 Mme1 Mme1 digestion and end polish using T4 polymerase AA XX NN TT 20bp Self-ligate , propagate in bacteria Extract single- PET plasmid DNA STEP8 18bp GIS single PET library AA M/B/S

17. STEP9 release and purifry 50-bp PET b M b Concanate PETs into vectore STEP10 GIS PET library

18. STEP11 Sequencing and mapping GIS PETs to genome Gen1 Gen2 Gen3

19.

20. PET transcriptome Genome annotation   

21.  

Pet

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