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Team:TU-Eindhoven/Protocols
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| - | <p>The principal protocol for yeast transformation was Gietz and Schiestl (2007)<html><a href="#ref_gietz_2007" name="text_gietz_2007"><sup>[1]</sup></a></html>. It promised high transformation efficiency, however, in combination with our yeast the yield was a factor thousand lower than expected. The solution was to increase the amount of DNA used for transformation up to a whopping 3 ug and to work with freshly cultured yeast in mid-log phase instead of vials of frozen competent cells. | + | <p>The principal protocol for yeast transformation was Gietz and Schiestl (2007)<html><a href="#ref_gietz_2007" name="text_gietz_2007"><sup>[1]</sup></a></html>. It promised high transformation efficiency, however, in combination with our yeast the yield was a factor thousand lower than expected. The solution was to increase the amount of DNA used for transformation up to a whopping 3 ug and to work with freshly cultured yeast in mid-log phase instead of vials of frozen competent cells. For more details, please refer to the [[File:yeast_transformation.pdf|modified yeast transformation protocol]].</p> |
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Revision as of 13:58, 25 September 2012
Standard protocols and modifications
In the lab we used many standard protocols, however, some of were modified from literature. The reasons for modification are indicated briefly below.
BioBrick construction
Yeast transformation
The principal protocol for yeast transformation was Gietz and Schiestl (2007)[1]. It promised high transformation efficiency, however, in combination with our yeast the yield was a factor thousand lower than expected. The solution was to increase the amount of DNA used for transformation up to a whopping 3 ug and to work with freshly cultured yeast in mid-log phase instead of vials of frozen competent cells. For more details, please refer to the File:Yeast transformation.pdf.
References
- [1]Gietz and Schiestl, ''High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method'', Nature Protocols (2007) vol. 2, issue 1, pp. 31-34
