Wiki/Team:Warsaw/protocols

From 2009.igem.org

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(New page: {{WarHead}} ==Lab protocols== Here is the place for the all protocols used by the Warsaw Team during the iGEM 2009. Feel free to add yours. Please be sure to use the <pre> {{Anchor|na...)
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==Lab protocols==
==Lab protocols==
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__TOC__
Here is the place for the all protocols used by the Warsaw Team during the iGEM 2009. Feel free to add yours.  
Here is the place for the all protocols used by the Warsaw Team during the iGEM 2009. Feel free to add yours.  
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Add desired volume of DNA to the 100-μl-culture in eppendorf tube. Incubate 30 min on ice. Heat shock for 90 s at 42&deg;C. Incubate 10 min on ice. Add 0.9 ml of culture medium and let the bacteria grow at 37&deg;C.  
Add desired volume of DNA to the 100-μl-culture in eppendorf tube. Incubate 30 min on ice. Heat shock for 90 s at 42&deg;C. Incubate 10 min on ice. Add 0.9 ml of culture medium and let the bacteria grow at 37&deg;C.  
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===Plasmid DNA isolation{{Anchor|miniprep}}===
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We use "Plasmid Mini" plasmid DNA isolation kit from A&A Biotechnology and follow the protocol of producer.
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===DNA isolation from agarose gel {{Anchor|gelout}}===
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We use "Gel-Out" DNA isolation kit from A&A Biotechnology and follow the protocol of producer.
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===DNA purification after enzymatic reaction {{Anchor|cleanup}}
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We use "Clean-Up" DNA purification kit from A&A Biotechnology and follow the protocol of producer.
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===Genomic DNA isolation {{Anchor|genomic_mini}}
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We use "Genomic-Mini" universal genomic DNA isolation kit from A&A Biotechnology and follow the protocol of producer.
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===DNA digest {{Anchor|dna_digest}}===
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We use restriction enzymes and buffers provided by Fermentas. Overall volume of digest mix is either 20 μl, either 50 μl in case of digesting for ligation. We usually use 1 μl of restriction enzyme and the buffer in 10x dilution (as they initially are 10x concentrated). The rest of mix is plasmid DNA.

Revision as of 11:29, 12 July 2009

Notebook Team Project Home

Lab protocols

Contents


Here is the place for the all protocols used by the Warsaw Team during the iGEM 2009. Feel free to add yours.

Please be sure to use the 

{{Anchor|name of protocol}}

tag before your desired protocol - it simplifies further linking to the protocols in notebook entries (then you will use just 

[https://2009.igem.org/Team:Warsaw/protocols#protocol_name protocol name]

to link to your protocol.

A great example of describing protocols you can find on the iGEM 2008 Warsaw Team page

Example:


Chemotransformation

Add desired volume of DNA to the 100-μl-culture in eppendorf tube. Incubate 30 min on ice. Heat shock for 90 s at 42°C. Incubate 10 min on ice. Add 0.9 ml of culture medium and let the bacteria grow at 37°C.

Plasmid DNA isolation

We use "Plasmid Mini" plasmid DNA isolation kit from A&A Biotechnology and follow the protocol of producer.

DNA isolation from agarose gel

We use "Gel-Out" DNA isolation kit from A&A Biotechnology and follow the protocol of producer.

===DNA purification after enzymatic reaction

We use "Clean-Up" DNA purification kit from A&A Biotechnology and follow the protocol of producer.

===Genomic DNA isolation

We use "Genomic-Mini" universal genomic DNA isolation kit from A&A Biotechnology and follow the protocol of producer.

DNA digest

We use restriction enzymes and buffers provided by Fermentas. Overall volume of digest mix is either 20 μl, either 50 μl in case of digesting for ligation. We usually use 1 μl of restriction enzyme and the buffer in 10x dilution (as they initially are 10x concentrated). The rest of mix is plasmid DNA.


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