Noi/NOTES/2011-5-28

From ZhangLabWiki
Jump to navigation Jump to search

BSPP DMR330k capture of HIV patient samples and N37[edit]

  • May 19, 2011, received 39 gDNA from the blood samples of HIV patients form Tropher's lab, Medical School, UCSD
  • Ma7 25, 2011, received 10 gDNA from all tissues of N37 donor from Jin Billy Li, Standford Univ
Sample ID PID New order in Zhang lab Place in 96plate Volume (ul) Conc. (ng/ul) Total amount (ug)
3 26406 102 1 A1 21 100 2.1
4 26406 108 2 A2 21 100 2.1
5 26604 054 3 A3 21 100 2.1
6 26604 066 4 A4 21 100 2.1
11 NW578 000 5 A5 21 100 2.1
12 NW578 012 6 A6 21 100 2.1
13 58346 006 7 A7 21 100 2.1
14 58346 012 8 A8 21 100 2.1
15 CA198 006 9 A9 21 100 2.1
16 CA198 C12 10 A10 21 100 2.1
17 CA219 000 11 A11 21 100 2.1
18 CA219 006 12 A12 21 100 2.1
19 CA275 000 13 B1 21 100 2.1
20 CA275 006 14 B2 21 100 2.1
21 CB252 000 15 B3 21 100 2.1
22 CB252 006 16 B4 21 100 2.1
24 CC182 012 17 B5 21 100 2.1
25 24371 000 18 B6 21 100 2.1
26 24371 012 19 B7 21 100 2.1
27 NW470 012 20 B8 21 100 2.1
28 NW470 024 21 B9 21 100 2.1
30 27054 012 22 B10 21 100 2.1
31 CA239 000 23 B11 21 100 2.1
33 CE214 000 24 B12 21 100 2.1
34 CE214 006 25 C1 21 100 2.1
35 CB255 000 26 C2 21 100 2.1
36 CB255 006 27 C3 21 100 2.1
37 21003 000 28 C4 21 100 2.1
38 21003 012 29 C5 21 100 2.1
39 CG139 006 30 C6 21 100 2.1
40 CG139 018 31 C7 21 100 2.1
41 NM633 012 32 C8 21 100 2.1
42 NM633 024 33 C9 21 100 2.1
43 NM805 012 34 C10 21 100 2.1
44 NM805 024 35 C11 21 100 2.1
45 CB199 000 36 C12 21 100 2.1
46 26592 060 37 D1 21 100 2.1
47 24054 030 38 D2 21 100 2.1
48 24054 036 39 D3 21 100 2.1


Number Sample Concentration (ug/ul) Volume (ul) Total amount (ug) NewID TE buffer (ul) Total volume (ul) Final conc. (ng/ul)
1.00 Cerebellum 1.38 1.45 2.00 N37-1 18.55 20.00 100.05
2.00 Colon 1.44 1.39 2.00 N37-2 18.61 20.00 100.08
3.00 Frontal lobe 0.91 2.20 2.00 N37-3 17.80 20.00 100.10
4.00 Heart 0.73 2.74 2.00 N37-4 17.26 20.00 100.01
5.00 Small intestine 1.09 1.83 1.99 N37-5 18.17 20.00 99.74
6.00 Liver 0.93 2.15 2.00 N37-6 17.85 20.00 99.98
7.00 Lung 0.50 4.00 2.00 N37-7 16.00 20.00 100.00
8.00 Skeletal muscle 0.57 3.51 2.00 N37-8 16.49 20.00 100.04
9.00 Pancrease 1.48 1.35 2.00 N37-9 18.65 20.00 99.90
10.00 Stomach 1.17 1.71 2.00 N37-10 18.29 20.00 100.04
  • Note: For N37 sample set, TE buffer was added to bring up volume to 20ul and adjusted the final concentration to ~100ng/ul

Bisulfite conversion[edit]

  • EZ-96 DNA Methylation-Gold Kit
  • Catalog# D5007 (Shallow-Well Format)
  • Reagent list

- CT conversion reagent: to the CT conversion reagent bottle, add 9mL H2O, 500ul M-dissolving buffer, and 3mL M-dilution buffer, mix at RT with frequent vortexing or shaking for 15min
- M-Binding buffer
- M-Wash buffer: add 144mL of 100% EtOH to the 36mL M-Wash buffer (total volume = 180mL)
- M-desulphonation buffer

  • Procedure
  1. calculate 1.2ug vol. of DNA sample, and the vol. of H2O to adjust total volume to 20ul
  2. add water to 96-conversion plate, then add DNA
  3. add 130ul of CT conversion reagent to 20ul of DNA sample, mix sample by pipetting up and down
  4. seal the plate with sticky film
  5. Transfer the plate to a thermal cycler and perform the following steps.
    1. 98C for 10mim
    2. 64C for 2.5hr
    3. 4 hold (can be up to 20hr)
  6. add 400ul of M-binding buffer to the well of the Silicon-A Binding plate mounted on a collection plate
  7. transfer the samples from the conversion plate to the well of the Silicon-A Binding plate, mix by pipetting up and down
  8. centrifuge at >3000xg for 5min, discard the flow through
  9. add 400ul of M-Wash buffer to each well, centrifuge at >3000xg for 5 min
  10. add 200ul of M-Desulphonation buffer to each well, incubate at RT (20C-30C) for 20min
  11. centrifuge at >3000xg for 5min, discard the flow through
  12. add 400ul of M-Wash buffer to each well, centrifuge at >3000xg for 5min, discard the flow through
  13. add another 400ul of M-Wash buffer, centrifuge for 10 min
  14. place the Silicon-A binding plate onto an Elution plate, add 15ul of M-Elution buffer directly to each well
  15. incubate for 5min, centrifuge at >3000xg for 3min to elute the DNA
  16. DNA samples are ready for the next analysis or store at -20C for later use
  • Note: for long term storage, store at or below -70C
  • Actual speed for centrifugation: 2,000rpm.
  • Note: The ID used for HIV samples will correspond to the position in the 96-well plate.
Order Sample ID in Zhang lab Conc. (ng/ul) volume for 1200ng (ul) H20 (ul) Total amount for bis-cvt (ng) CT conversion reagent (ul) Total volume (ul)
1 A1 100 12 8 1200 130 150
2 A2 100 12 8 1200 130 150
3 A3 100 12 8 1200 130 150
4 A4 100 12 8 1200 130 150
5 A5 100 12 8 1200 130 150
6 A6 100 12 8 1200 130 150
7 A7 100 12 8 1200 130 150
8 A8 100 12 8 1200 130 150
9 A9 100 12 8 1200 130 150
10 A10 100 12 8 1200 130 150
11 A11 100 12 8 1200 130 150
12 A12 100 12 8 1200 130 150
13 B1 100 12 8 1200 130 150
14 B2 100 12 8 1200 130 150
15 B3 100 12 8 1200 130 150
16 B4 100 12 8 1200 130 150
17 B5 100 12 8 1200 130 150
18 B6 100 12 8 1200 130 150
19 B7 100 12 8 1200 130 150
20 B8 100 12 8 1200 130 150
21 B9 100 12 8 1200 130 150
22 B10 100 12 8 1200 130 150
23 B11 100 12 8 1200 130 150
24 B12 100 12 8 1200 130 150
25 C1 100 12 8 1200 130 150
26 C2 100 12 8 1200 130 150
27 C3 100 12 8 1200 130 150
28 C4 100 12 8 1200 130 150
29 C5 100 12 8 1200 130 150
30 C6 100 12 8 1200 130 150
31 C7 100 12 8 1200 130 150
32 C8 100 12 8 1200 130 150
33 C9 100 12 8 1200 130 150
34 C10 100 12 8 1200 130 150
35 C11 100 12 8 1200 130 150
36 C12 100 12 8 1200 130 150
37 D1 100 12 8 1200 130 150
38 D2 100 12 8 1200 130 150
39 D3 100 12 8 1200 130 150
40 N37-1 100 12 8 1200 130 150
41 N37-2 100 12 8 1200 130 150
42 N37-3 100 12 8 1200 130 150
43 N37-4 100 12 8 1200 130 150
44 N37-5 100 12 8 1200 130 150
45 N37-6 100 12 8 1200 130 150
46 N37-7 100 12 8 1200 130 150
47 N37-8 100 12 8 1200 130 150
48 N37-9 100 12 8 1200 130 150
49 N37-10 100 12 8 1200 130 150


  • Plate layout
Plat layout 1 2 3 4 5 6 7 8 9 10 11 12
A A A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12
B
C B B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12
D
E C C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12
F
G D D1 D2 D3
H N37-1 N37-2 N37-3 N37-4 N37-5 N37-6 N37-7 N37-8 N37-9 N37-10


  • 2011_06_01

Qubit ss-DNA assay[edit]

  • added 1ul of bis-cvt gDNA in total volume 200ul, so dilution factor = 200)
sample ID Conc. In Qubit (ng/mL) Final conc. (ng/ul) Volume (ul) Yields (ng)
A1 184.00 36.80 15.00 552.00
A2 316.00 63.20 15.00 948.00
A3 208.00 41.60 15.00 624.00
A4 224.00 44.80 15.00 672.00
A5 176.00 35.20 15.00 528.00
A6 218.00 43.60 15.00 654.00
A7 240.00 48.00 15.00 720.00
A8 220.00 44.00 15.00 660.00
A9 365.00 73.00 15.00 1095.00
A10 220.00 44.00 15.00 660.00
A11 216.00 43.20 15.00 648.00
A12 285.00 57.00 15.00 855.00
B1 191.00 38.20 15.00 573.00
B2 263.00 52.60 15.00 789.00
B3 198.00 39.60 15.00 594.00
B4 153.00 30.60 15.00 459.00
B5 234.00 46.80 15.00 702.00
B6 301.00 60.20 15.00 903.00
B7 148.00 29.60 15.00 444.00
B8 209.00 41.80 15.00 627.00
B9 215.00 43.00 15.00 645.00
B10 193.00 38.60 15.00 579.00
B11 168.00 33.60 15.00 504.00
B12 240.00 48.00 15.00 720.00
C1 169.00 33.80 15.00 507.00
C2 230.00 46.00 15.00 690.00
C3 248.00 49.60 15.00 744.00
C4 204.00 40.80 15.00 612.00
C5 208.00 41.60 15.00 624.00
C6 139.00 27.80 15.00 417.00
C7 127.00 25.40 15.00 381.00
C8 174.00 34.80 15.00 522.00
C9 215.00 43.00 15.00 645.00
C10 202.00 40.40 15.00 606.00
C11 200.00 40.00 15.00 600.00
C12 179.00 35.80 15.00 537.00
D1 231.00 46.20 15.00 693.00
D2 234.00 46.80 15.00 702.00
D3 213.00 42.60 15.00 639.00
N37-1 201.00 40.20 15.00 603.00
N37-2 135.00 27.00 15.00 405.00
N37-3 194.00 38.80 15.00 582.00
N37-4 195.00 39.00 15.00 585.00
N37-5 165.00 33.00 15.00 495.00
N37-6 282.00 56.40 15.00 846.00
N37-7 206.00 41.20 15.00 618.00
N37-8 187.00 37.40 15.00 561.00
N37-9 256.00 51.20 15.00 768.00
N37-10 168.00 33.60 15.00 504.00


Capture set up[edit]

Probe calculation[edit]

Subset ID Adjusted Multiplier based on qPCR (Multiplier E) Actual amount of probe required (probe:target ratio, 100:1) Multiplier E x amount probe required Probe concentration (ng/ul) Volume (ul) Volume for 55rxn (ul)
DMR.s1 0.05 2.23 0.11 1.86 0.06 3.30
DMR.s2 0.07 2.23 0.16 1.95 0.08 4.57
DMR.s3 0.23 18.90 4.41 23.20 0.19 10.44
DMR.s4 1.00 18.90 18.90 23.20 0.81 44.81
DMR.s5 4.83 18.90 91.29 29.20 3.13 171.94
DMR.Exp1 0.23 18.90 4.44 31.88 0.14 7.65
DMR.Exp2 0.19 18.90 3.50 24.11 0.15 8.00
DMR.Exp3 0.27 18.90 5.03 28.90 0.17 9.56
105nM Oligo suppressor 1.00 55.00
Total volume 5.73 315.28


Reaction mix[edit]

Components 1rxn (ul) 55rxn mix (ul)
10X AmpLigase Buffer 2.50 137.50
DMR330k probe mix (ul) 4.73 260.15
105nM Oligo suppressor 1.00 55.00
Total volume (ul) 8.23 452.65


sample ID Final conc. (ng/ul) Volume for 400ng(ul) H2O (ul) Total amount of bis-cvt gDNA (ng) Probe mix (ul) 10x Amligase buffer (ul) 105nM Oligo suppressor (ul) Total volume (ul)
A1 36.80 10.87 5.90 400.00 4.73 2.50 1.00 25.00
A2 63.20 6.33 10.44 400.00 4.73 2.50 1.00 25.00
A3 41.60 9.62 7.15 400.00 4.73 2.50 1.00 25.00
A4 44.80 8.93 7.84 400.00 4.73 2.50 1.00 25.00
A5 35.20 11.36 5.41 400.00 4.73 2.50 1.00 25.00
A6 43.60 9.17 7.60 400.00 4.73 2.50 1.00 25.00
A7 48.00 8.33 8.44 400.00 4.73 2.50 1.00 25.00
A8 44.00 9.09 7.68 400.00 4.73 2.50 1.00 25.00
A9 73.00 5.48 11.29 400.00 4.73 2.50 1.00 25.00
A10 44.00 9.09 7.68 400.00 4.73 2.50 1.00 25.00
A11 43.20 9.26 7.51 400.00 4.73 2.50 1.00 25.00
A12 57.00 7.02 9.75 400.00 4.73 2.50 1.00 25.00
B1 38.20 10.47 6.30 400.00 4.73 2.50 1.00 25.00
B2 52.60 7.60 9.17 400.00 4.73 2.50 1.00 25.00
B3 39.60 10.10 6.67 400.00 4.73 2.50 1.00 25.00
B4 30.60 13.07 3.70 400.00 4.73 2.50 1.00 25.00
B5 46.80 8.55 8.22 400.00 4.73 2.50 1.00 25.00
B6 60.20 6.64 10.13 400.00 4.73 2.50 1.00 25.00
B7 29.60 13.51 3.26 400.00 4.73 2.50 1.00 25.00
B8 41.80 9.57 7.20 400.00 4.73 2.50 1.00 25.00
B9 43.00 9.30 7.47 400.00 4.73 2.50 1.00 25.00
B10 38.60 10.36 6.41 400.00 4.73 2.50 1.00 25.00
B11 33.60 11.90 4.87 400.00 4.73 2.50 1.00 25.00
B12 48.00 8.33 8.44 400.00 4.73 2.50 1.00 25.00
C1 33.80 11.83 4.94 400.00 4.73 2.50 1.00 25.00
C2 46.00 8.70 8.07 400.00 4.73 2.50 1.00 25.00
C3 49.60 8.06 8.71 400.00 4.73 2.50 1.00 25.00
C4 40.80 9.80 6.97 400.00 4.73 2.50 1.00 25.00
C5 41.60 9.62 7.15 400.00 4.73 2.50 1.00 25.00
C6 27.80 14.39 2.38 400.00 4.73 2.50 1.00 25.00
C7 25.40 15.75 1.02 400.00 4.73 2.50 1.00 25.00
C8 34.80 11.49 5.28 400.00 4.73 2.50 1.00 25.00
C9 43.00 9.30 7.47 400.00 4.73 2.50 1.00 25.00
C10 40.40 9.90 6.87 400.00 4.73 2.50 1.00 25.00
C11 40.00 10.00 6.77 400.00 4.73 2.50 1.00 25.00
C12 35.80 11.17 5.60 400.00 4.73 2.50 1.00 25.00
D1 46.20 8.66 8.11 400.00 4.73 2.50 1.00 25.00
D2 46.80 8.55 8.22 400.00 4.73 2.50 1.00 25.00
D3 42.60 9.39 7.38 400.00 4.73 2.50 1.00 25.00
N37-1 40.20 9.95 6.82 400.00 4.73 2.50 1.00 25.00
N37-2 27.00 14.81 1.96 400.00 4.73 2.50 1.00 25.00
N37-3 38.80 10.31 6.46 400.00 4.73 2.50 1.00 25.00
N37-4 39.00 10.26 6.51 400.00 4.73 2.50 1.00 25.00
N37-5 33.00 12.12 4.65 400.00 4.73 2.50 1.00 25.00
N37-6 56.40 7.09 9.68 400.00 4.73 2.50 1.00 25.00
N37-7 41.20 9.71 7.06 400.00 4.73 2.50 1.00 25.00
N37-8 37.40 10.70 6.07 400.00 4.73 2.50 1.00 25.00
N37-9 51.20 7.81 8.96 400.00 4.73 2.50 1.00 25.00
N37-10 33.60 11.90 4.87 400.00 4.73 2.50 1.00 25.00


Program

   -> 95c 30sec -> cool down to 55C at 0.02C/sec -> 55C 20h 
   -> add 2.5ul SLN mix (2U/ul AmpliTaq Stoffel fragment; 0.5U/ul AmpLigase; 100uM dNTP) (2011_06_02)
   -> 55C 20h-> 94C 2min -> add 2ul Exo I/III mix-> 37C 2h -> 94C 2min -> 4C hold. (2011_06_03)

SLN mix[edit]

Components Stock conc. Unit Final conc. Unit Prepare volume 150
AmpliTaq Stoffel 10.00 U/ul 2.00 U/ul 30.00
AmpLigase 5.00 U/ul 0.50 U/ul 15.00
dNTP 1.00 mM 100.00 uM 15.00
10x AmpLigase Buffer 10.00 x 1.00 x 15.00
H2O 75.00
Total 150.00


  • aliquot 15ul of SLN to strip tube, and add 2.5 ul to each tube.
  • Note:
    • There was something wrong with sample in well A9-A12. The volume of reaction in these four well was lower than 10ul, but before capture the levels of the reaction in every well were the same. I think that it's possible that the 96-well plate was not sealed very well especially the border of the plate, so the evaporation possibly occurred. There was proof that the reagents were not missing added to these four wells since one of them could be amplified, but not in a good yield. In the future, there should be more concerns about the plate sealing.
    • It would be better if the samples will be rearranged from A1-H1, ..., A2-H2,.... rather than A1-A12, B1-B12. This because the multichannel pipette has 8 channels, so it would be easier to add samples in the same column at once. In addition, it's less possibility to miss adding reagents in well No.9-12

PCR Amplification with barcoded primers (AmpF6.4Sol and AmpR6.3 IndX)[edit]

Components 1rxn 52 rxn mix
Captured template 8.00 0.00
10uM AmpF6.4Sol 2.00 104.00
10uM AmpR6.3 (IndX, X 1,2,3,…,49) 2.00 0.00
50x SYBG 0.80 41.60
2x Phusion MM 50.00 2600.00
H2O 37.20 1934.40
Total 100.00 Aliquot 90ul, add 8ul template and 2 ul of AmpR6.3 IndX


Plate layout

A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12
AmpR6.3 Ind1 AmpR6.3 Ind2 AmpR6.3 Ind3 AmpR6.3 Ind4 AmpR6.3 Ind5 AmpR6.3 Ind6 AmpR6.3 Ind7 AmpR6.3 Ind8 AmpR6.3 Ind9 AmpR6.3 Ind10 AmpR6.3 Ind11 AmpR6.3 Ind12
B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12
AmpR6.3 Ind13 AmpR6.3 Ind14 AmpR6.3 Ind15 AmpR6.3 Ind16 AmpR6.3 Ind17 AmpR6.3 Ind18 AmpR6.3 Ind19 AmpR6.3 Ind20 AmpR6.3 Ind21 AmpR6.3 Ind22 AmpR6.3 Ind23 AmpR6.3 Ind24
C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12
AmpR6.3 Ind25 AmpR6.3 Ind26 AmpR6.3 Ind27 AmpR6.3 Ind28 AmpR6.3 Ind29 AmpR6.3 Ind30 AmpR6.3 Ind31 AmpR6.3 Ind32 AmpR6.3 Ind33 AmpR6.3 Ind34 AmpR6.3 Ind35 AmpR6.3 Ind36
D1 D2 D3 NTC
AmpR6.3 Ind37 AmpR6.3 Ind38 AmpR6.3 Ind39 AmpR6.3 Ind49
N37-1 N37-2 N37-3 N37-4 N37-5 N37-6 N37-7 N37-8 N37-9 N37-10
AmpR6.3 Ind40 AmpR6.3 Ind41 AmpR6.3 Ind42 AmpR6.3 Ind43 AmpR6.3 Ind44 AmpR6.3 Ind45 AmpR6.3 Ind46 AmpR6.3 Ind47 AmpR6.3 Ind48 AmpR6.3 Ind49


Program
98C 30s -> (98C 10s -> 58C 20s -> 72C 20s)x5 -> (98C 10s -> 72C 20s)x13 -> 72C 3min

Real-time PCR curves[edit]

File:A1-A8.png File:B1-B12.png File:C1-C12.png File:D1-D3.png File:N37-1-10 NTC.png
2011_06_06

  • Amplify one more reaction for each sample excluded A9-A12 needed to be repeated for capturing
  • Increase DNA captured template from 8ul to 10ul
Components 1rxn 47 rxn mix
Captured template 10.00 0.00
10uM AmpF6.4Sol 2.00 94.00
10uM AmpR6.3 (IndX, X 1,2,3,…,49) 2.00 0.00
50x SYBG 0.80 37.60
2x Phusion MM 50.00 2350.00
H2O 35.20 1654.40
Total 100.00 Aliquot 88ul, add 10ul template and 2ul AmpR6.3IndX


A1 A2 A3 A4 A5 A6 A7 A8 ' ' ' '
AmpR6.3 Ind1 AmpR6.3 Ind2 AmpR6.3 Ind3 AmpR6.3 Ind4 AmpR6.3 Ind5 AmpR6.3 Ind6 AmpR6.3 Ind7 AmpR6.3 Ind8
B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12
AmpR6.3 Ind13 AmpR6.3 Ind14 AmpR6.3 Ind15 AmpR6.3 Ind16 AmpR6.3 Ind17 AmpR6.3 Ind18 AmpR6.3 Ind19 AmpR6.3 Ind20 AmpR6.3 Ind21 AmpR6.3 Ind22 AmpR6.3 Ind23 AmpR6.3 Ind24
C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12
AmpR6.3 Ind25 AmpR6.3 Ind26 AmpR6.3 Ind27 AmpR6.3 Ind28 AmpR6.3 Ind29 AmpR6.3 Ind30 AmpR6.3 Ind31 AmpR6.3 Ind32 AmpR6.3 Ind33 AmpR6.3 Ind34 AmpR6.3 Ind35 AmpR6.3 Ind36
D1 D2 D3
AmpR6.3 Ind37 AmpR6.3 Ind38 AmpR6.3 Ind39
N37-1 N37-2 N37-3 N37-4 N37-5 N37-6 N37-7 N37-8 N37-9 N37-10
AmpR6.3 Ind40 AmpR6.3 Ind41 AmpR6.3 Ind42 AmpR6.3 Ind43 AmpR6.3 Ind44 AmpR6.3 Ind45 AmpR6.3 Ind46 AmpR6.3 Ind47 AmpR6.3 Ind48 AmpR6.3 Ind49


File:PCR-round2 2011 06 06.png

Ct Values[edit]

  • Calculated from first round and second round real-time PCR
Sample ID Average Ct Stdev
A1 14.99 NA
A2 13.86 0.45
A3 15.01 0.09
A4 14.78 0.07
A5 13.42 0.78
A6 13.92 0.11
A7 14.58 0.04
A8 14.80 0.00
B1 13.17 0.52
B2 14.64 0.16
B3 15.23 0.11
B4 15.33 0.23
B5 15.37 0.21
B6 14.30 0.06
B7 15.81 0.11
B8 15.04 0.34
B9 15.31 0.44
B10 15.65 0.33
B11 16.04 0.42
B12 13.99 0.20
C1 12.81 0.37
C2 14.20 0.07
C3 14.62 0.08
C4 13.00 0.30
C5 15.46 0.11
C6 16.69 0.11
C7 16.89 0.32
C8 14.84 0.13
C9 15.66 0.33
C10 15.70 0.06
C11 16.05 0.30
C12 16.08 0.02
D1 15.47 0.57
D2 15.66 0.25
D3 15.53 0.45
N37-1 14.46 0.06
N37-2 15.26 0.23
N37-3 13.36 0.51
N37-4 14.89 0.02
N37-5 14.21 0.02
N37-6 15.74 0.20
N37-7 14.99 0.13
N37-8 15.60 0.08
N37-9 15.99 0.65
N37-10 15.40 1.06


  • Continued on 2011_06_06: [[1]]