Luckau Protocols: Difference between revisions

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==Tara Luckau Protocols==
{|{{table}} width="900"
Clark Laboratory
|-
| width="600" style="background-color: #BCED91;" align="center"| <span style="font-size:32px;"> Tara's Protocols </span>


===Agarose Gel===
|align="right" style="background-color: #9DB68C;" |
====Purpose====
<span style="font-size:16px;"> [[User:Tara_K._Luckau | Tara K. Luckau's Home Page]] </span>
Agarose gels are used to verify the presence and estimate the size of DNA (both genomic and amplified). When loaded into a gel and subjected to an electric current, DNA fragments migrate to the positive terminal. Small DNA fragments migrate more rapidly than large ones. By running a commercially available ladder of fragments of known size, we can estimate the DNA fragment size in our sample of interest.


To visualize the DNA after electrophoresis, a dye called Gel Red is added during the agarose-making process. The dye binds to the DNA and flouresces when exposed to UV light.
[[User:Tara_K._Luckau/Notebook/Team_ConGen | Conservation Genetics Lab Notebook]]
====Materials To Be Familiar With====
* Gel Rig, caster tray, combs
* [[Image:OwlA2Large.jpg]]  [[Image:OwlA2Large_draw.jpg|300 px]]


====Protocol====
[[Luckau_Protocols | Tara's Protocols]]
# Make gel using 1x TAE and agarose
#* Use 1% for genomic DNA, 2% for amplified DNA
{| border="1" style="margin: 1em auto 1em auto" "text-align: center"
|-
! Gel Rig Size !! 1x TAE (mL) !! 1.5% agarose (g) !! 2% agarose (g) !! Gel Red (µL) !! Sample volume (µL) !! max voltage (V)
|-
| small || 50 || _ || 1 || 5 || 4-6 || 80
|-
| rowspan="2"| medium || rowspan="2"| 130 || rowspan="2"| _ || rowspan="2"| 2.6 || rowspan="2"| 13 || 24-tooth comb: 6-10 || rowspan="2"| 120
|-
| 36-tooth comb: 3-5
|-
| large || _ || _ || _ || _ || _ || 170
|}
|}
#
 
 
==Protocols used in Dr. Rulon Clark's laboratory==
 
 
===Lab Processes===
 
 
* [[Luckau_Protocols:NanoDrop | NanoDrop]]
* [[Luckau_Protocols:PCR | PCR]]
* [[Luckau_Protocols:Agarose Gel | Agarose Gel]]
* [[Luckau_Protocols:FragmentAnalysisSubmission | Fragment Analysis Submission]]
* [[Luckau_Protocols:ShipDryIce | Shipping with Dry Ice]]
 
 
===Reagents===
 
 
* [[Luckau_Protocols:Low TE | Low TE]]
* [[Luckau_Protocols:Tris-Cl | Tris-Cl]]
* [[Luckau_Protocols:KCl | KCl]]
* [[Luckau_Protocols:MgCl2 | MgCl<sub>2</sub>]]
* [[Luckau_Protocols:TAE | TAE]]
* [[Luckau_Protocols:1Kb Ladder | 1Kb Ladder]]
* [[Luckau_Protocols:dNTPs | dNTPs]]
* [[Luckau_Protocols:PCR Buffers A-H | PCR Buffers A-H]]
* [[Luckau_Protocols:PrimerResuspension | Primer Resuspension]]
 
 
===Software Programs===
 
 
* [[Luckau_Protocols:GeneMarker | GeneMarker]]
* [[Luckau_Protocols:Scoring | How I Score Frag Data]]
* [[Luckau_Protocols:STRUCTURE | STRUCTURE]]
 
 
===Inventory===
 
 
* [[Luckau_Protocols:Inventory | Inventory & Catalog #s]]

Latest revision as of 15:05, 29 May 2013

Tara's Protocols

Tara K. Luckau's Home Page

Conservation Genetics Lab Notebook

Tara's Protocols


Protocols used in Dr. Rulon Clark's laboratory

Lab Processes


Reagents


Software Programs


Inventory

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