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Wiki/Team:Warsaw/protocols
From 2009.igem.org
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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°C. Incubate 10 min on ice. Add 0.9 ml of culture medium and let the bacteria grow at 37°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°C. Incubate 10 min on ice. Add 0.9 ml of culture medium and let the bacteria grow at 37°C. | ||
| - | ===Plasmid DNA isolation{{Anchor|miniprep}}=== | + | ===Plasmid DNA isolation {{Anchor|miniprep}}=== |
We use "Plasmid Mini" plasmid DNA isolation kit from A&A Biotechnology and follow the protocol of producer. | We use "Plasmid Mini" plasmid DNA isolation kit from A&A Biotechnology and follow the protocol of producer. | ||
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We use "Gel-Out" DNA isolation kit from A&A Biotechnology and follow the protocol of producer. | We use "Gel-Out" DNA isolation kit from A&A Biotechnology and follow the protocol of producer. | ||
| - | ===DNA purification after enzymatic reaction {{Anchor|cleanup}} | + | ===DNA purification after enzymatic reaction {{Anchor|cleanup}}=== |
We use "Clean-Up" DNA purification kit from A&A Biotechnology and follow the protocol of producer. | We use "Clean-Up" DNA purification kit from A&A Biotechnology and follow the protocol of producer. | ||
| - | ===Genomic DNA isolation {{Anchor|genomic_mini}} | + | ===Genomic DNA isolation {{Anchor|genomic_mini}}=== |
We use "Genomic-Mini" universal genomic DNA isolation kit from A&A Biotechnology and follow the protocol of producer. | We use "Genomic-Mini" universal genomic DNA isolation kit from A&A Biotechnology and follow the protocol of producer. | ||
Revision as of 11:30, 12 July 2009
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Lab protocols
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:
ChemotransformationAdd 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 isolationWe use "Plasmid Mini" plasmid DNA isolation kit from A&A Biotechnology and follow the protocol of producer. DNA isolation from agarose gelWe use "Gel-Out" DNA isolation kit from A&A Biotechnology and follow the protocol of producer. DNA purification after enzymatic reactionWe use "Clean-Up" DNA purification kit from A&A Biotechnology and follow the protocol of producer. Genomic DNA isolationWe use "Genomic-Mini" universal genomic DNA isolation kit from A&A Biotechnology and follow the protocol of producer. DNA digestWe 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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