Difference between revisions of "Team:Oxford/Notebook/Week1"
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Revision as of 12:20, 2 July 2015
22/06/2015
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22/06/2015
| Date | Researcher(s) | Content | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 22/06/2015 | Whole Team |
Preparation of Stock Solutions1. gBlocksThe gBlocks ordered from IDT arrived in the form of vials of 200ng solid DNA powder. (refer to BioBricks page for information on DNA sequences) The gBlocks were made into 10ng/µl stock solutions in Milli-Q water for storage:
2. PrimersThe forward and reverse primers ordered from IDT came in 32.4nmol and 34.3nmol of solid respectively. (Sequences: Forward - CTTTTTTGCCGGACTGC; Reverse - ATGATTTCTGGAATTCGC) The primers were made into 100µM stock solutions in Milli-Q water for storage:
Preparation of Reaction Solutions 1. gBlocks2µl of each stock solution were diluted in Milli-Q water to achieve final solution volumes of 20µl to make 1ng/µl-1 reaction solutions. 2. Primers2µl of each stock solution were diluted in Milli-Q water to achieve final solution volumes of 20µl to make 10µM reaction solutions. (These solutions are labelled as “Prefix primer” and “suffix primer” in eppendorf tubes in the fridge) Polymerase Chain Reaction Set-upThe protocol for running a PCR using NEB’s Q5 High-Fidelity 2X Master Mix can be found here. Contact us: E-mail: oxfordigem@bioch.ox.ac.uk | Mailing Address: Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom
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