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Team:Reading/Protocols
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| - | Prior to PCR, it was ensured that all DNA obtained from miniprep was of a concentration of at least 1ng/ul per 100bp; this was performed using a desktop Thermo Scientific NanoDrop machine. All PCR tubes were kept on ice prior to usage. | + | Prior to PCR, it was ensured that all DNA obtained from miniprep was of a concentration of at least 1ng/ul per 100bp; this was performed using a desktop Thermo Scientific NanoDrop machine. All PCR tubes were kept on ice prior to usage. |
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| + | 2μl of each of the VFR (forward) and VR (reverse) primers were added to sterile PCR tubes, along with 2μl of each respective transformed plasmid and 14μl mastermix [add reference here?]. | ||
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Revision as of 15:18, 13 October 2014
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A note on protocols |
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Below are a number of examples of protocols - modified and non-modified - used throughout our project. The protocols we list below may not have been followed exactly in each instance of usage, often due to time constraints - beginning an experiment on one day and concluding it on another. |
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The Goods
Here are all of the protocols.
PCR
PCR was used as a means of amplifying each one of our biobrick constructs. Each construct, along with its corresponding flanking sequence of 50-100 bases, was amplified out of each transformed pSB1C3 plasmid.
Prior to PCR, it was ensured that all DNA obtained from miniprep was of a concentration of at least 1ng/ul per 100bp; this was performed using a desktop Thermo Scientific NanoDrop machine. All PCR tubes were kept on ice prior to usage.
2μl of each of the VFR (forward) and VR (reverse) primers were added to sterile PCR tubes, along with 2μl of each respective transformed plasmid and 14μl mastermix [add reference here?].
Potassium ferricyanide assay
Another potential protocol
E. coli transformation
Another potential protocol
Synechocystis transformation
Another potential protocol
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ReferencesHere are the references. |
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AcknowledgementsEveryone we need to thank for help with protocols. |
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rusynbioigem@gmail.com |
