1. Analysis of transcription related H3/H4 displacement at the individual gene level

2. 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 Experiments

3. Showed change in AltB expression as expected Showed possible enrichment of PolIIon AltB in late G2 Previously... Late G2 Mid G2 Imp UnIP Imp Un IP

4. 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 Experiments  

5. Mapping of AltB -230 -156

6. Comparison of mononucleosomes from control and flag incorporated cells Testing effect of Flag histone incorporation

7. Determine relative amplification from DNA cut 5 min and 15 min compared to uncut Testing Mnase sensitivity of AltB

8. AltBnucleosomepostition in Mid (off) and Late (on) G2 -230 -156 100bp

9. Confirmation of quantitative mapping -230 -156 Primer extension assay

10. Development of a novel method for high resolution mapping -230 -156 Tried to quantify/amplify of different products in primer extension reaction Late G2 Mid G2 PE Q1 Q2 Q3 Q4 Q5 PE Q1 Q2 Q3 Q4 Q5

11. Single primer tests -230 -156 Neat 1:100 Neat 1:100 Mid G2 Late G2 P1/P2 P1 P2 P1/P2 P1 P2 P1/P2 P1 P2 P1/P2 P1 P2

12. Use reconstituted 5s nucleosomes Confirm position restriction digest New method Apply new method to map completely AltB Validation of new mapping method