Noi/NOTES/2012-8-22

From ZhangLabWiki
Jump to navigation Jump to search
  • [Link to calendar]
  • Received 16 cell pellet (1x10^6 cells/tube) samples from Guanghui
No. Sample description Mark on the tube Lab ID
1 WT-ESC H9-WT-ES GL-1
2 LRRK2-GS-ESC H9-LRRK-ES GL-2
3 WT-NSCs H9-WT-NSC-P9 GL-3
4 LRRK2-GS-NSCs H9-LRRK-NSC-P9 GL-4
5 2nd-iPSCs-WT#5 (early passage) H9-WT-NSC-iPSC#5 P18 GL-5
6 2nd-iPSCs-WT#3 (early passage) H9-WT-NSC-iPSC#3 P18 GL-6
7 2nd-iPSCs-LK2-GS#2 (early passage) H9-LRRK-NSC-iPSC#2 P18 GL-7
8 2nd-iPSCs-LK2-GS#6 (early passage) H9-LRRK-NSC-iPSC#6 P18 GL-8
9 2nd-iPSCs-WT#5 (late passage) W5-ips P34 GL-9
10 2nd-iPSCs-WT#3 (late passage) W2-ips P34 GL-10
11 2nd-iPSCs-LK2-G2#2 (late passage) L2-ips P34 GL-11
12 2nd-iPSCs-LK2-GS#6 (late passage) L6-ips P34 GL-12
13 Cardiomyocyes combine two tube to get 1M cells GL-13
14 WT-NSCs FA52-#6-NSC GL-14
15 FA-NSCs FA123#2-NSC GL-15
16 Corrected FA-NSCs FA-GC-16-NEO-NSC GL-16
  • Sample code GL1-GL13 for Project 1
  • Sample code GL14-GL16 for Project 2
  • Extract gDNA 16 of GL and 3 of AL
GL= Eluted with AE buffer, volume 55ul
AL= Eluted with AE buffer, volume 200ul	
  • Sample Info will be added later

gDNA concetration by Nanodrop

Sample ID ng/ul A260 260/280 Yields (ug)
GL-1 295.8 5.916 2.04 16.3
GL-2 220.6 4.412 2.07 12.1
GL-3 149.7 2.994 2.01 8.2
GL-4 172.3 3.445 2.06 9.5
GL-5 285.2 5.704 2.05 15.7
GL-6 296.6 5.932 2.03 16.3
GL-7 320.0 6.400 2.03 17.6
GL-8 339.3 6.786 2.04 18.7
GL-9 467.5 9.349 2.02 25.7
GL-10 431.7 8.633 2.01 23.7
GL-11 349.1 6.981 2.03 19.2
GL-12 234.5 4.691 2.04 12.9
GL-13 72.2 1.445 1.89 4.0
GL-14 137.3 2.747 2.01 7.6
GL-15 207.5 4.150 2.03 11.4
GL-16 135.5 2.710 2.00 7.5
AL-3 256.8 5.136 2.08 51.4
AL-4 142.1 2.842 2.04 28.4
AL-5 140.1 2.802 2.00 28.0


Bisulfite conversion[edit]

Lab IDs Conc. (ng/ul) Volume for 1200ng LabID ID H2O Total amount (ng) CT conversion reagent (ul) Total volume (ul)
GL-1 295.8 4.06 GL-1 15.94 1200.00 130.00 150.00
GL-2 220.6 5.44 GL-2 14.56 1200.00 130.00 150.00
GL-3 149.7 8.02 GL-3 11.98 1200.00 130.00 150.00
GL-4 172.3 6.97 GL-4 13.03 1200.00 130.00 150.00
GL-5 285.2 4.21 GL-5 15.79 1200.00 130.00 150.00
GL-6 296.6 4.05 GL-6 15.95 1200.00 130.00 150.00
GL-7 320.0 3.75 GL-7 16.25 1200.00 130.00 150.00
GL-8 339.3 3.54 GL-8 16.46 1200.00 130.00 150.00
GL-9 467.5 2.57 GL-9 17.43 1200.00 130.00 150.00
GL-10 431.7 2.78 GL-10 17.22 1200.00 130.00 150.00
GL-11 349.1 3.44 GL-11 16.56 1200.00 130.00 150.00
GL-12 234.5 5.12 GL-12 14.88 1200.00 130.00 150.00
GL-13 72.2 16.61 GL-13 3.39 1200.00 130.00 150.00
GL-14 137.3 8.74 GL-14 11.26 1200.00 130.00 150.00
GL-15 207.5 5.78 GL-15 14.22 1200.00 130.00 150.00
GL-16 135.5 8.86 GL-16 11.14 1200.00 130.00 150.00
AL-1 226.6 5.30 AL-1 14.70 1200.00 130.00 150.00
AL-2 375.5 3.20 AL-2 16.80 1200.00 130.00 150.00
AL-3 256.8 4.67 AL-3 15.33 1200.00 130.00 150.00
AL-4 142.1 8.45 AL-4 11.55 1200.00 130.00 150.00
AL-5 140.1 8.56 AL-5 11.44 1200.00 130.00 150.00
Program
1. 98°C for 8 minutes
2. 54°C for 60 minutes
3. 4°C hold