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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) | ||
==== 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. We got no bands from the 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). | |||
=====Reagents:===== | |||
* O. indoliflex gDNA (H): 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 1ul gDNA (concentration TBA), 6.5ul dH20. | |||
* O. indoliflex gDNA (UCL): 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 1ul gDNA (concentration TBA), 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 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 3 from O. indoliflex: 12.5ul Taq readymix, 2.5ul FP, 2.5ul RP, 2ul DNA, 5.5ul dH20. | |||
* PSBIC3 (plasmid backbone - used as positive control): 12.5ul Taq readymix, 1ul FP, 1ul RP, 3ul DNA, 7.5ul dH20. | |||
=====PCR:===== | |||
'''''We do not know whether this program was ran as it is written below, but here are the instructions''''' | |||
* Initial D: 95C 30s | |||
* '''30 cycles of:''' | |||
* 95C 15s | |||
* 55C 30s | |||
* 68C 50s | |||
* Final extension 65C 5 mins | |||
* Hold 4C ∞ | |||
=====Visualisation:===== | |||
[[File:Gel_run_15_9.jpg|left|400px|thumb|none|Gel run 15th sept]] | |||
'''From left:''' | |||
*1: 250-10000bp ladder (from hackspace). Ladder [http://www.taq-dna.com/rich_files/attachments/DNA_Ladders_DNA_Weight_markers_DNA_leiter/1000_bp_1kb_DNA_Ladder.pdf key] | |||
*2: O. indofilex template from hackspace PCR product. | |||
*3: O. indofilex template from UCL PCR product. | |||
*4: ANF 1 PCR product. | |||
*5: ANF 2 PCR product. | |||
*6: ANF 3 PCR product. | |||
*7: PSBIC3 PCR product. | |||
