Daniel:Notebook/HiResChrPaint/2013-6-10: Difference between revisions

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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]]
|}


[[Daniel:Notebook/HiResChrPaint|Back to Calendar]]
===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


Volume Report: [[File:Dan_GelQuant_061013.xls]]
|-  align="center" valign="bottom"
|style="font-weight:bold" height="12" | V4S2*
| align="center" | 158.7
| align="center" | 1.6


[[Image:GelQuant_061013.jpg|600px]]
|-  align="center" valign="bottom"
|style="font-weight:bold" height="12" | V6S2*
| align="center" | 153.4
| align="center" | 1.5


[[Image:GelQuant_051513_Volume.jpg]]
|}
Note: *denotes no aa-dUTP

Latest revision as of 22:53, 2 July 2013

Probe Prep 3 (Started 5/29/2013)[edit]

Back to Calendar

TBU Gel Quantification[edit]

Protocol 2

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