Gene of interest
1
Gene of interest Unique Sequence TTAATTAA PacI NotI Target sequence (Sense) Target sequence (Antisense) 9bp loop Complimentary Oligo Mix 10 µg of each oligo and heat at 65°C for 20 minutes and incubate at 22°C for 1 hour Digested overnight with 20 units each of Pac1 and Not1 enzyme Ligate into Pac1 and Not1 restricted γ genome of BSMV-VIGS vector Hairpin-encoding VIGS construct Antisense VIGS construct Figure S2 GCGGCCGC TTCAAGAGA Sense Oligo 45 bp 45 bp AATTAATT CGCCGGCG AAGTTCTCT TTAATTAA PacI NotI Target sequence (Antisense) GCGGCCGC Sense Oligo 90 bp Complimentary Oligo AATTAATT CGCCGGCG TAA GC TTCAAGAGA TAATT CGCCGG AAGTTCTCT TAA GC TAATT CGCCGG 5’ 3’ 3’ 5’ 5’ 3’ 3’ 5’ 5’ 3’ 5’ 3’ 5’ 3’ 5’ 3’
description
Gene of interest. Unique Sequence. Sense Oligo. Sense Oligo. Target sequence (Antisense). Target sequence (Antisense). Target sequence (Sense). PacI. NotI. PacI. 9bp loop. NotI. GCGGCCGC. 3’. TTAATTAA. 5’. 5’. TTAATTAA. TTCAAGAGA. GCGGCCGC. 3’. 45 bp. 45 bp. 90 bp. 3’. - PowerPoint PPT Presentation
Transcript of Gene of interest
Gene of interest
Unique Sequence
TTAATTAA
PacI NotITarget sequence
(Sense)Target sequence
(Antisense)9bp loop
Complimentary Oligo
Mix 10 µg of each oligo and heat at 65°C for 20 minutes and incubate at 22°C for 1 hour
Digested overnight with 20 units each of Pac1 and Not1 enzyme
Ligate into Pac1 and Not1 restricted γ genome of BSMV-VIGS vector
Hairpin-encoding VIGS construct
Antisense VIGS construct
Figure S2
GCGGCCGCTTCAAGAGA
Sense Oligo
45 bp45 bpAATTAATT CGCCGGCGAAGTTCTCT
TTAATTAA
PacI NotITarget sequence
(Antisense)
GCGGCCGC
Sense Oligo
90 bp
Complimentary Oligo
AATTAATT CGCCGGCG
TAA GCTTCAAGAGATAATT CGCCGGAAGTTCTCT
TAA GCTAATT CGCCGG
5’
3’
3’
5’
5’
3’
3’
5’
5’3’
5’3’5’
3’5’3’
