Team:Northwestern/Protocol

From 2010.igem.org

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<!--- The Mission, Experiments --->
<!--- The Mission, Experiments --->
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{|- style="font-size: 87%; background: #AFC7C7; color: #000000" cellpadding="1" cellspacing="1" bordercolor="#000000" width="75%" align="center"
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{|- style="font-size: 87%; background: #EFCDF8; color: #FFFFFF" cellpadding="1" cellspacing="1" bordercolor="#000000" width="75%" align="center"
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!align="center"|[[Team:Northwestern|<font color="#FFFFFF">'''Home'''</font>]]
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!align="center"|[[Team:Northwestern|<font color="#2B3856">'''Home'''</font>]]
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!align="center"|[[Team:Northwestern/Brainstorm|<font color="#FFFFFF">'''Brainstorm'''</font>]]
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!align="center"|[[Team:Northwestern/Brainstorm|<font color="#000000">'''Brainstorm'''</font>]]
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!align="center"|[[Team:Northwestern/Team|<font color="#FFFFFF">'''Team'''</font>]]
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!align="center"|[[Team:Northwestern/Team|<font color="#000000">'''Team'''</font>]]
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!align="center"|[[Team:Northwestern/Acknowledgements|<font color="#FFFFFF">'''Acknowledgements'''</font>]]
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!align="center"|[[Team:Northwestern/Acknowledgements|<font color="#000000">'''Acknowledgements'''</font>]]
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!align="center"|[[Team:Northwestern/Project|<font color="#FFFFFF">'''Project'''</font>]]
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!align="center"|[[Team:Northwestern/Project|<font color="#000000">'''Project'''</font>]]
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!align="center"|[[Team:Northwestern/SideProject|<font color="#FFFFFF">'''Human Practices'''</font>]]
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!align="center"|[[Team:Northwestern/SideProject|<font color="#000000">'''Human Practices'''</font>]]
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!align="center"|[[Team:Northwestern/Parts|<font color="#FFFFFF">'''Parts'''</font>]]
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!align="center"|[[Team:Northwestern/Parts|<font color="#000000">'''Parts'''</font>]]
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!align="center"|[[Team:Northwestern/Notebook|<font color="#FFFFFF">'''Notebook'''</font>]]
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!align="center"|[[Team:Northwestern/Notebook|<font color="#000000">'''Notebook'''</font>]]
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!align="center"|[[Team:Northwestern/Calendar|<font color="#FFFFFF">'''Calendar'''</font>]]
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!align="center"|[[Team:Northwestern/Calendar|<font color="#000000">'''Calendar'''</font>]]
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!align="center"|[[Team:Northwestern/Protocol|<font color="#2B3856">'''Protocol'''</font>]]
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!align="center"|[[Team:Northwestern/Protocol|<font color="#000000">'''Protocol'''</font>]]
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!align="center"|[[Team:Northwestern/Safety|<font color="#FFFFFF">'''Safety'''</font>]]
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!align="center"|[[Team:Northwestern/Safety|<font color="#000000">'''Safety'''</font>]]
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!align="center"|[[Team:Northwestern/Links|<font color="#FFFFFF">'''Links'''</font>]]
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!align="center"|[[Team:Northwestern/Links|<font color="#000000">'''Links'''</font>]]
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!align="center"|[[Team:Northwestern/References|<font color="#FFFFFF">'''References'''</font>]]
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!align="center"|[[Team:Northwestern/References|<font color="#000000">'''References'''</font>]]
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!align="center"|[[Team:Northwestern/Media|<font color="#FFFFFF">'''Media'''</font>]]
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!align="center"|[[Team:Northwestern/Media|<font color="#000000">'''Media'''</font>]]
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!align="center"|[[Team:Northwestern/Contact|<font color="#FFFFFF">'''Contact'''</font>]]
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!align="center"|[[Team:Northwestern/Contact|<font color="#000000">'''Contact'''</font>]]
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{| align="center" border="0" width="75%"
{| align="center" border="0" width="75%"
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=='''Protocols'''==
== '''DNA''' ==
== '''DNA''' ==
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[[Team:Northwestern/Protocol/Fluorescence Microscopy|Fluorescence Microscopy]]
[[Team:Northwestern/Protocol/Fluorescence Microscopy|Fluorescence Microscopy]]
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=='''Reagents'''==
=='''Reagents'''==
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[[Team:Northwestern/Protocol/Reagents|Reagents]]
[[Team:Northwestern/Protocol/Reagents|Reagents]]
=='''Cell Staining'''==
=='''Cell Staining'''==
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[[Team:Northwestern/Protocol/Rhodamine-Conjugated Chitin Probe|Rhodamine-Conjugated Chitin Probe]]
[[Team:Northwestern/Protocol/Rhodamine-Conjugated Chitin Probe|Rhodamine-Conjugated Chitin Probe]]
[[Team:Northwestern/Protocol/Methanol Fixation|Methanol Fixation]]
[[Team:Northwestern/Protocol/Methanol Fixation|Methanol Fixation]]
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==Methanol Fixation==
 
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[[Team:Northwestern/Protocol/LIVE/DEAD® BacLight - Bacterial Viability Kit|Live/Dead Baclight]]
[[Team:Northwestern/Protocol/LIVE/DEAD® BacLight - Bacterial Viability Kit|Live/Dead Baclight]]
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===[[Prepping DNA from the kit plates]]===
 
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===[[Quikchange (from primers to colonies!)]]===
 
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===[[Kit to Stock Plasmid]]===
 
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===[[Ethanol Precipitation]]===
 
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===[[New Part Design(PCR)]]===
 
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=='''Bacterial Work'''==
 
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===[[Transformation]]===
 
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===[[O/N Culture]]===
 
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===[[Preparation of Competent Cells]]===
 
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===[[LB Media]]===
 
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===[[Preparing Plates]]===
 
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===[[Glycerol Stocks]]===
 
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=='''Assembly'''==
 
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===[[Restriction Enzyme Digests]]===
 
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===[[Plasmid Construction]]===
 
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===[[3A Assembly]]===
 
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===[[Ligations]]===
 
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===[[Mini Prep]]===
 
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=='''Microscopy'''==
 
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===[[Confocal Microscopy]]===
 
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===[[Fluorescence Microscopy]]===
 
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=='''Reagents'''==
 
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===[[Reagents]]===
 
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=='''Cell Staining'''==
 
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===[[Rhodamine-Conjugated Chitin Probe]]===
 
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==Methanol Fixation==
 
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# Rinse slide with ethanol and flame
 
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# Rinse slide with PBS and dry with KimWipe
 
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# Apply cell sample to microscope slide and let air dry
 
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# Submerge in -20C absolute methanol for 5-10 min
 
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# Wash 3 times with 1X TBS (submerge in 1X TBS for 5 minutes per wash)
 
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** '''Do not let cells dry for the rest of the procedure'''
 
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===[[LIVE/DEAD® BacLight - Bacterial Viability Kit]]===
 
|}
|}

Latest revision as of 01:23, 27 October 2010


Tech Institute
Home Brainstorm Team Acknowledgements Project Human Practices Parts Notebook Calendar Protocol Safety Links References Media Contact

Protocols

DNA

Prepping DNA from the kit plates

Quikchange: Primers to Colonies

Kit to Stock Plasmid

Ethanol Precipitation

New Part Design

Bacterial Work

Transformation

Overnight Cultures

Preparation of Competent Cells

LB Media

Preparing Plates

Glycerol Stocks

Assembly

Restriction Enzyme Digests

Plasmid Construction

3A Assembly

Ligations

Qiagen Mini Prep

Microscopy

Confocal Microscopy

Fluorescence Microscopy

Reagents

Reagents

Cell Staining

Rhodamine-Conjugated Chitin Probe

Methanol Fixation

Live/Dead Baclight

Retrieved from "http://2010.igem.org/Team:Northwestern/Protocol"