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Project:Public Biobrick: Difference between revisions
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*4: O. Indofilex gDNA sample 1 (band just visible) | *4: O. Indofilex gDNA sample 1 (band just visible) | ||
*5: O. Indofilex gDNA sample 1 (band just visible) | *5: O. Indofilex gDNA sample 1 (band just visible) | ||
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==== 14-15/9: Hackspace ==== | ==== 14-15/9: Hackspace ==== | ||
Yeping, Simon and Tonderai prepared and PCRed the following 25ul volume reactions on the 14/9, Will visualised them on the 15/9 | Yeping, Simon and Tonderai prepared and PCRed the following 25ul volume reactions on the 14/9, Will visualised them on the 15/9. | ||
=====Reagents:===== | =====Reagents:===== | ||
* O. indoliflex gDNA (H): 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 1ul gDNA (concentration | * O. indoliflex gDNA (H): 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 1ul gDNA (1:100 concentration), 6.5ul dH20. | ||
* O. indoliflex gDNA (UCL): 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 1ul gDNA (concentration | * O. indoliflex gDNA (UCL): 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 1ul gDNA (1:100 concentration), 6.5ul dH20. | ||
* ANF template 1 from O. indoliflex: 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 2ul DNA, 5.5ul dH20. | * ANF template 1 from O. indoliflex: 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 2ul DNA, 5.5ul dH20. | ||
* ANF template 2 from O. indoliflex: 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP (primers diluted 1:2), 2ul DNA, 5.5ul dH20. | * ANF template 2 from O. indoliflex: 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP (primers diluted 1:2), 2ul DNA, 5.5ul dH20. | ||
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=====PCR:===== | =====PCR:===== | ||
'''''We do not know whether this program was ran as it is written below, but here are the instructions''''' | '''''We do not know whether this program was actually ran as it is written below, but here are the instructions.''''' | ||
* Initial D: 95C 30s | * Initial D: 95C 30s | ||
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* Hold 4C ∞ | * Hold 4C ∞ | ||
=====Visualisation:===== | =====Visualisation and interpretation:===== | ||
We electrophoresed on a 1% agarose gel for 40 mins. | |||
[[File:Gel_run_15_9.jpg|left|400px|thumb|none|Gel run 15th sept]] | [[File:Gel_run_15_9.jpg|left|400px|thumb|none|Gel run 15th sept]] | ||
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*7: PSBIC3 PCR product. | *7: PSBIC3 PCR product. | ||
So we got no bands from the O. indofilex gDNA reactions, but we did get bands at the right position (285 bp) from 2/3 of the ANF template ones. (Not sure yet whether this means PCR succeeded, as 'ANF template' may already have been this size. Will explain what ANF template means when I understand it better myself!). We also got a band from the positive control at the right position (2070bp). | |||
