Daniel:Notebook/ComboLock/2016-10-13: Difference between revisions
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<li>Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A-C by reusing the water</li> | <li>Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A-C by reusing the water</li> | ||
</ol></ol> | </ol></ol> | ||
=Padlock0301 Dilution Test= | |||
===Stock Dilutions=== | |||
Create the following dilutions of PCC Template by taking 10 uL stock and diluting with 90 uL nfH2O (note:zero is normal 10 uM stock). #6 is the highest concentration, made on [[Daniel:Notebook/ComboLock/2016-10-7|Friday 10-7]]. | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="background-color:#8DB4E2;font-size:12pt;font-weight:bold" align="center" | |||
| width="65" height="30" | Tube No | |||
| width="65" | Dilution | |||
| width="65" | Molecules Available | |||
| width="65" | Amount | |||
|- style="font-size:12pt" align="center" | |||
| align="center" height="15" | 6 | |||
| align="center" | 1.00E+06 | |||
| align="center" | 1.00E+06 | |||
| 10 amol | |||
|- style="background-color:#BFBFBF;font-size:12pt" align="center" | |||
| align="center" height="15" | 7 | |||
| align="center" | 1.00E+07 | |||
| align="center" | 1.00E+05 | |||
| 1 amol | |||
|- style="font-size:12pt" align="center" | |||
| align="center" height="15" | 8 | |||
| align="center" | 1.00E+08 | |||
| align="center" | 1.00E+04 | |||
| 100 zmol | |||
|- style="background-color:#BFBFBF;font-size:12pt" align="center" | |||
| align="center" height="15" | 9 | |||
| align="center" | 1.00E+09 | |||
| align="center" | 1000 | |||
| 10 zmol | |||
|- style="font-size:12pt" align="center" | |||
| align="center" height="15" | 10 | |||
| align="center" | 1.00E+10 | |||
| align="center" | 100 | |||
| 1 zmol | |||
|- style="background-color:#BFBFBF;font-size:12pt" align="center" | |||
| align="center" height="15" | 11 | |||
| align="center" | 1.00E+11 | |||
| align="center" | 10 | |||
| 100 ymol | |||
|- style="font-size:12pt" align="center" | |||
| align="center" height="15" | 12 | |||
| align="center" | 1.00E+12 | |||
| align="center" | 1 | |||
| 10 ymol | |||
|} | |||
===Buffers=== | |||
==Protocol== | |||
[[Category:ComboLock]] [[Category:20161007]] | [[Category:ComboLock]] [[Category:20161007]] |
Revision as of 18:09, 13 October 2016
Padlock0301 Test (Started Friday 10-7)
Library Prep
- Size Select Gel
- Centrifuge tubes at 12000 rpm for 1.5 minutes to bring gel to bottom
- Extract supernatant and add to nanosep column
- Centrifuge at 12000 rpm for 1.5 minutes; collect supernatant
- Ethanol Precipitation
- Add 1250 uL 100% EtOH, 50 uL 3M NaOAc, and 1.5 uL glycoblue to each sample
- Incubate at -80C for a while
- Centrifuge for 25 minutes at 12000 rpm at 4C
- Remove supernatant and add 750 uL 70% EtOH
- Centrifuge for 12 minutes at 12000 rpm at 4C
- Remove supernatant and spin in vacuum centrifuge for 10 minutes
- Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A-C by reusing the water
Padlock0301 Dilution Test
Stock Dilutions
Create the following dilutions of PCC Template by taking 10 uL stock and diluting with 90 uL nfH2O (note:zero is normal 10 uM stock). #6 is the highest concentration, made on Friday 10-7.
Tube No | Dilution | Molecules Available | Amount |
6 | 1.00E+06 | 1.00E+06 | 10 amol |
7 | 1.00E+07 | 1.00E+05 | 1 amol |
8 | 1.00E+08 | 1.00E+04 | 100 zmol |
9 | 1.00E+09 | 1000 | 10 zmol |
10 | 1.00E+10 | 100 | 1 zmol |
11 | 1.00E+11 | 10 | 100 ymol |
12 | 1.00E+12 | 1 | 10 ymol |