Daniel:Notebook/HiResChrPaint/2013-6-10: Difference between revisions
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>B1lake (Created page with " Back to Calendar Volume Report: File:Dan_GelQuant_061013.xls 600px Image:GelQuant_051513_Volume.jpg") |
>Djacobse |
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=Probe Prep 3 ([[Daniel:Notebook/HiResChrPaint/2013-5-29|Started 5/29/2013]])= | |||
[[Daniel:Notebook/HiResChrPaint|Back to Calendar]] | |||
==TBU Gel Quantification== | |||
[[Daniel:Protocols/TBUgel|Protocol 2]] | |||
1. Sample matrix | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
|style="font-weight:bold" width="65" height="15" | Lane | |||
| align="center" width="65" | 2 | |||
| align="center" width="65" | 3 | |||
| align="center" width="65" | 4 | |||
| align="center" width="65" | 5 | |||
| align="center" width="65" | 6 | |||
| align="center" width="65" | 7 | |||
| align="center" width="65" | 8 | |||
| align="center" width="65" | 9 | |||
| align="center" width="65" | 10 | |||
| align="center" width="65" | 11 | |||
|- style="font-size:12pt;font-weight:bold" align="center" valign="bottom" | |||
| height="30" | Sample | |||
| Low Mass Ladder | |||
| Low Mass Ladder | |||
| V4S2 | |||
| V6S2 | |||
| V4S2* | |||
| V6S2* | |||
| V4S2 | |||
| V6S2 | |||
| V4S2* | |||
| V6S2* | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="15" | uL sample | |||
| align="center" | 0.5 | |||
| align="center" | 1 | |||
| align="center" | 0.5 | |||
| align="center" | 0.5 | |||
| align="center" | 0.5 | |||
| align="center" | 0.5 | |||
| 2** | |||
| 2** | |||
| 2** | |||
| 2** | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="15" | uL TBE | |||
| align="center" | 1.5 | |||
| align="center" | 1 | |||
| align="center" | 9.5 | |||
| align="center" | 9.5 | |||
| align="center" | 9.5 | |||
| align="center" | 9.5 | |||
| align="center" | 1 | |||
| align="center" | 1 | |||
| align="center" | 1 | |||
| align="center" | 1 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="30" | uL 2X Urea Buffer | |||
| align="center" | 2 | |||
| align="center" | 2 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 1 | |||
| align="center" | 1 | |||
| align="center" | 1 | |||
| align="center" | 1 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="15" | Dilution | |||
| NA | |||
| NA | |||
| align="center" | 1:10 | |||
| align="center" | 1:10 | |||
| align="center" | 1:10 | |||
| align="center" | 1:10 | |||
| align="center" | 1:20 | |||
| align="center" | 1:20 | |||
| align="center" | 1:20 | |||
| align="center" | 1:20 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="15" | Loaded | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
| align="center" | 4 | |||
|} | |||
2. Ran gel for 18 minutes at 250 V | |||
3. Stained for 25 minutes with 3 uL SYBR Gold | |||
===Results=== | |||
Volume Report: [[File:GelQuant_061013.xls]] | |||
{| class="wikitable" | |||
|- | |||
| [[Image:GelQuant_061013.jpg|400px]] || [[Image:GelQuant_051513_Volume.jpg|400px|Volumes used for quantification]] | |||
|} | |||
[[ | ===Nanodrop Results=== | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-weight:bold" align="center" valign="bottom" | |||
| width="69" height="12" | Sample | |||
| width="69" | ng/uL | |||
| width="69" | ug in 10 uL | |||
|- align="center" valign="bottom" | |||
|style="font-weight:bold" height="12" | V4S2 | |||
| align="center" | 119.9 | |||
| align="center" | 1.2 | |||
|- align="center" valign="bottom" | |||
|style="font-weight:bold" height="12" | V6S2 | |||
| align="center" | 122.9 | |||
| align="center" | 1.2 | |||
|- align="center" valign="bottom" | |||
|style="font-weight:bold" height="12" | V4S2* | |||
| align="center" | 158.7 | |||
| align="center" | 1.6 | |||
|- align="center" valign="bottom" | |||
|style="font-weight:bold" height="12" | V6S2* | |||
| align="center" | 153.4 | |||
| align="center" | 1.5 | |||
|} | |||
Note: *denotes no aa-dUTP |
Latest revision as of 22:53, 2 July 2013
Probe Prep 3 (Started 5/29/2013)[edit]
TBU Gel Quantification[edit]
1. Sample matrix
Lane | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 |
Sample | Low Mass Ladder | Low Mass Ladder | V4S2 | V6S2 | V4S2* | V6S2* | V4S2 | V6S2 | V4S2* | V6S2* |
uL sample | 0.5 | 1 | 0.5 | 0.5 | 0.5 | 0.5 | 2** | 2** | 2** | 2** |
uL TBE | 1.5 | 1 | 9.5 | 9.5 | 9.5 | 9.5 | 1 | 1 | 1 | 1 |
uL 2X Urea Buffer | 2 | 2 | 10 | 10 | 10 | 10 | 1 | 1 | 1 | 1 |
Dilution | NA | NA | 1:10 | 1:10 | 1:10 | 1:10 | 1:20 | 1:20 | 1:20 | 1:20 |
Loaded | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 |
2. Ran gel for 18 minutes at 250 V 3. Stained for 25 minutes with 3 uL SYBR Gold
Results[edit]
Volume Report: File:GelQuant 061013.xls
File:GelQuant 061013.jpg | Volumes used for quantification |
Nanodrop Results[edit]
Sample | ng/uL | ug in 10 uL |
V4S2 | 119.9 | 1.2 |
V6S2 | 122.9 | 1.2 |
V4S2* | 158.7 | 1.6 |
V6S2* | 153.4 | 1.5 |
Note: *denotes no aa-dUTP