Giles 23 April 09
-
Upload
gilesoelliott -
Category
Documents
-
view
131 -
download
0
Transcript of Giles 23 April 09
![Page 1: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/1.jpg)
![Page 2: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/2.jpg)
Following to be done for ArdC, ProP and AltB at
mid and late G2
◦ Quantification of transcription
◦ Quantification of PolII occupancy
◦ Quantification of H3/H4 replacement
Perform high resolution mapping at mid and late
G2 for AltB
![Page 3: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/3.jpg)
Showed change in
AltB expression as
expected
Showed possible
enrichment of PolII on
AltB in late G2
0
1000
2000
3000
4000
5000
6000
ArdC ProP AltB
Mid G2
Late G2
-20
0
20
40
60
80
100
120
140
ArdC ProP AltB
Mid G2
Late G2
Late G2 Mid G2
Imp Un IP Imp Un IP
![Page 4: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/4.jpg)
Following to be done for ArdC, ProP and AltB at
mid and late G2
◦ Quantification of transcription
◦ Quantification of PolII occupancy
◦ Quantification of H3/H4 replacement
Perform high resolution mapping at mid and late
G2 for AltB
![Page 5: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/5.jpg)
-230 -156
![Page 6: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/6.jpg)
Comparison of mononucleosomes from control and flag incorporated cells
-8
-6
-4
-2
0
2
4
6
8
10
1 2 3 4 5 6 6 7
8a
8b 9
10
11a
11B
12
13
14
15
16
17
18
19
20
Ard
C
Pro
P
DC
t
MC-C MF-F
![Page 7: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/7.jpg)
Determine relative amplification from DNA cut 5 min and 15 min compared to uncut
0
5
10
15
20
25
1 2 3 4 5 6 7 8A 8B 9 10 11A 11B 12 13 14 15 17 19 ArdC
dCt5 dCt15
![Page 8: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/8.jpg)
-4
-3
-2
-1
0
1
2
3
4
1 2 3 4 5 6 7 8a 8b 9 10 11a 11b 12 13 14 15 16 17 18 19 20
Mid G2 Late G2
100bp
-230 -156
![Page 9: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/9.jpg)
Primer extension assay
-230 -156
![Page 10: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/10.jpg)
Tried to quantify/amplify of different products in primer extension reaction
-230 -156
Mid G2 Late G2
PE Q1 Q2 Q3 Q4 Q5 PE Q1 Q2 Q3 Q4 Q5
![Page 11: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/11.jpg)
P1/P2 P1 P2P1/P2 P1 P2 P1/P2 P1 P2P1/P2 P1 P2
Neat 1:100 Neat 1:100
Mid G2 Late G2
-230 -156
![Page 12: Giles 23 April 09](https://reader030.fdocuments.in/reader030/viewer/2022020207/559f54261a28ab17778b458c/html5/thumbnails/12.jpg)
Use reconstituted 5s nucleosomes
Confirm position ◦ restriction digest
◦ New method
Apply new method to map completely AltB