Talk:20.109(S12):Aptamer binding assay (Day7)
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Binding Data Files
| Group | T/R zip data file | W/F zip data file |
|---|---|---|
| Red | Media:S12-M1-TR_Red.csv | Media:S12-M1-WF_Red.csv |
| Orange | Media:S12-M1-TR_Orange.csv | Media:S12-M1-WF_Orange.csv |
| Yellow | Media:S12-M1-TR_Yellow.csv | Media:S12-M1-WF_Yellow.csv |
| Green | Media:S12-M1-TR_Green.csv | Media:S12-M1-WF_Green.csv |
| Blue | Media:S12-M1-TR_Blue.csv | Media:S12-M1-WF_Blue.csv |
| Pink | Media:S12-M1-TR_Pink.csv | Media:S12-M1-WF_Pink.csv |
| Purple | Media:S12-M1-TR_Purple.csv | Media:S12-M1-WF_Purple.csv |
Binding Data Results Summary
T/R
* Be sure to post the values you got after baseline correction, not the raw data (if your curves didn't line up)!
* Order: heme, 6-5, 8-12, "pre" mix, post low wash, post high wash
* Recommended syntax for A405 column: "#1 ; #2; #3 <br> #4; #5; #6"
| Group | Corrected A405 values* | Pre % 8-12: expected, measured | Post % 8-12: fewer washes, more washes (measured) | Additional comments | |
|---|---|---|---|---|---|
| Syntax sample | 0.1; 0.2; 0.3 0.4; 0.5; 0.6 |
10%, 12% | 40%, 50% (4, 24 washes) | We used Grey group's 6-5 because we didn't have any. | |
| Pink | 0.14025053 (heme) 0.1422987 (6-5 pre) 0.21195409 (8-12 pre) 0.13888393 (mix pre) 0.16228816 (4 wash) 0.17831185 (24 wash) |
0.5%, -0.049% | 28.70% (4 wash), 51.70% (24 wash) | We started with a 0.5% 8-12 mixture and washed with 4 and 24 washes to measure enrichment | |
| Orange | 0.034495637(heme) 0.03498132(pre6-5) 0.045843026(pre8-12) 0.03623888(pre-mix) 0.041412034 (4 Wash) 0.040929238(24 Wash) |
1%, 11.5% | 59.2% (4 Washes), 54.7% (24 Washes) | We started with a 1% 8-12 mixture and washed with 4 and 24 washes to measure enrichment. We also generated very little RNA (~.3nmol), and thus had to dilute everything down to this amount. |
|
| Yellow | 0.155;0.160212;0.2070; 0.1642;0.1768;0.1776 |
0.25%, 8.6% | 35%, 37%(4 and 24 washes) | We started with 0.25% 8-12 mixture and washed with 4 and 24 washes | |
| Red | 0.218110 (heme) 0.211707 (6-5 pre) 0.277449 (8-12 pre) 0.215079 (mix pre) 0.237636 (4 wash) 0.256438 (24 wash) |
2% (expected) 5.13% (measured) |
39.44% (4 washes) 62.91% (24 washes) |
Started with 2% 8-12 mixture and washed with 4 and 24 washes. | |
| Green | 0.182316407(heme) 0.174965575(6-5 pre) 0.257898369(8-12 pre) 0.178721126 (mix pre) 0.185683139 (8 wash) 0.192153301(36 wash) |
1% (expected),4.52 %(measured) | 12.92, 20.72 % (8 and 36 washes) | Started with 1% 8-12 mixture and washed with 8 and 36 washes. | |
| Purple | 0.084126; 0.085602645; 0.13737727; 0.09304013; 0.14838624; 0.12533093 |
10% (expected), 14.37% (measured) | 121.1 % (4 washes), 76.7% (24 washes) | Started with 10% 8-12 mixture and washed with 4 and 24 washes. | |
| Blue | 0.0415141 (heme) 0.040754 (6-5 pre) 0.068389 (8-12 pre) 0.05368(mix pre) 0.066094(10 wash) 0.067595 (30 wash) |
10% (expected), 46.77% (measured) | 91.69% (10 washes), 97.13% (30 washes) | Started with 10% 8-12 mixture and washed with 10 and 30 washes. | |
W/F
* Be sure to post the values you got after baseline correction, not the raw data (if your curves didn't line up)!
* Order: heme, 6-5, 8-12, "pre" mix, post low wash, post high wash
* Recommended syntax for A405 column: "#1 ; #2; #3 <br> #4; #5; #6"
| Group | Corrected A405 values* | Pre % 8-12: expected, measured | Post % 8-12: fewer washes, more washes (measured) | Additional comments | |
|---|---|---|---|---|---|
| Syntax sample | 0.1; 0.2; 0.3 0.4; 0.5; 0.6 |
10%, 12% | 40%, 50% (4, 24 washes) | We used Grey group's 6-5 because we didn't have any. | |
| Purple | 0.116802; 0.114059; 0.189595 0.11671; 0.148567 (4degC); 0.161949 (22degC) |
2%, 3.51% | 45.89% (4degC), 63.39% (22degC) | We varied temperature (4deg, 22degC), keeping 20 washes constant. | |
| Orange | 0.172318067; 0.240679354; 0.174157415 0.188253912 (4 washes); 0.18944774(24 washes) |
0.5%, 2.69% | 23.31% (4 washes), 25.05% (24 washes) | We lowered the pre-selection % of 8-12 keeping average wash conditions of 4 and 24 washes. | |
| Blue | 0.139942214; 0.136271253; 0.220732734 0.147463577; 0.179008305 (8 Washes); 0.170598522 (16 Washes) |
10.0%, 14.0% | 53.4% (8 Washes), 42.9% (16 Washes) | ||
| Red | .1380; .1326; .1620 .1320; 1709 (12 washes); .1669 (18 washes); .1678 (24 washes) |
10%,negative percentage | more than 100% for all three wash conditions (12, 18, 24) |
| |
| Green | 0.101436131 (heme) 0.116573468 (6-5 pre) 0.175222094 (8-12 pre) 0.143155725 (mix pre) 0.141258046 (24 wash) |
50%, 44.26% | 41.1% (24 wash) | We lost our other sample (4 washes) due to a problem in either the wash step or the RTPCR step, so we had just one sample. | |
| Yellow | 0.1091; 0.1751; 0.2597 0.1728; 0.1868; 0.2064 |
1%, -2.6% (wrong) | 14% (4 washes), 37% (24 washes) | Something has definitely gone wrong! We aren't sure what yet, but you may want to exclude our data from your analysis :/ | |
| Pink | 0.16402881 (blank) 0.18001003(heme) 0.17570107 (pre 6-5) 0.22806994 (pre 8-12) 0.18219722 (pre mix) 0.19595873 (2 wash) 0.19819184 (34 wash) |
10%, 12.4% | 38.7% (2 wash), 42.9% (34 wash) | ||

