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Newcastle University Wetlab/11 August 2008
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(New page: {{:Team:Newcastle_University/WetLabCalendarHeader}} ==Monday 11th August== * Plasmids were isolated from pUc57-ncl08 (the ncl08 fragment contains the ''spaRK'' 2-part component syste...) |
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==Monday 11th August== | ==Monday 11th August== | ||
| - | * Plasmids were [[isolated]] from pUc57-ncl08 (the ncl08 fragment contains the ''spaRK'' 2-part component system). We aim to clone this into pGFP-rrnB and pJWV021 vectors. | + | * Plasmids were [[Isolating Plasmid from Cells (Miniprep)|isolated]] from pUc57-ncl08 (the ncl08 fragment contains the ''spaRK'' 2-part component system). We aim to clone this into pGFP-rrnB and pJWV021 vectors. |
* Restrictions of pGFP-rrnB and pJWV021 carried out using 10μl plasmid and 0.5μl in 50μl total reaction volume (see [[Restricting Plasmids (Double Restriction)]]). pJWV021 was restricted in a 2-step reaction as we beleived that the enzymes were incompatible in the same buffer. The second restriction of this plasmid used 48μl of the purified plasmid mixture in a total volume of 100μl. pGFP-rrnB was restricted using EcoRI and NHeI. pJWV021 was restricted firstly with NHeI and secondly with BglII. | * Restrictions of pGFP-rrnB and pJWV021 carried out using 10μl plasmid and 0.5μl in 50μl total reaction volume (see [[Restricting Plasmids (Double Restriction)]]). pJWV021 was restricted in a 2-step reaction as we beleived that the enzymes were incompatible in the same buffer. The second restriction of this plasmid used 48μl of the purified plasmid mixture in a total volume of 100μl. pGFP-rrnB was restricted using EcoRI and NHeI. pJWV021 was restricted firstly with NHeI and secondly with BglII. | ||
These were purified and stored overnight at -25˚C. | These were purified and stored overnight at -25˚C. | ||
Latest revision as of 16:29, 28 October 2008
Newcastle University
GOLD MEDAL WINNER 2008
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Monday 11th August
- Plasmids were isolated from pUc57-ncl08 (the ncl08 fragment contains the spaRK 2-part component system). We aim to clone this into pGFP-rrnB and pJWV021 vectors.
- Restrictions of pGFP-rrnB and pJWV021 carried out using 10μl plasmid and 0.5μl in 50μl total reaction volume (see Restricting Plasmids (Double Restriction)). pJWV021 was restricted in a 2-step reaction as we beleived that the enzymes were incompatible in the same buffer. The second restriction of this plasmid used 48μl of the purified plasmid mixture in a total volume of 100μl. pGFP-rrnB was restricted using EcoRI and NHeI. pJWV021 was restricted firstly with NHeI and secondly with BglII.
These were purified and stored overnight at -25˚C.
