Team:Goettingen/NoteBook w7

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July

15th

Analysis of the RD samples from 12.7

Analysis of the RD samples from 12.7.

Test run of plasmids purified on 12.7.13, vector and inserts on agarose gel

Samples:

Component	Part 7 plasmid (179.3 ng/µl)	pSB1C3 (2^nd elution, 3.4 ng/µl)	plasmid RBS-GFP-Terminator C12 (200.8 ng/µl)	plasmid RBS-GFP-Terminator C12 R.D.	DarR purified PCR product (4 µl primer; reaction from 24.6.13; 15.2 ng/µl)	DarR insert
DNA	0.5 µl	4 µl	0.5 µl	5 µl	2 µl	3 µl
dH₂O	3.5 µl	-	3.5 µl	-	2 µl	1 µl
5xLD	1 µl	1 µl	1 µl	-(sample in green FD buffer	1 µl	1 µl

Component	Riboswitch iGEM_40/41 Purified PCR product (45.7 ng/µl)	Riboswitch iGEM_40/41 insert	Riboswitch iGEM_41/42 Purified PCR product (36.8 ng/µl)	Riboswitch iGEM_41/42 insert	Riboswitch iGEM_40/43 Purified PCR product (47.7 ng/µl)	Riboswitch iGEM_40/43 insert	Riboswitch iGEM_42/43 Purified PCR product (35.7 ng/µl)	Riboswitch iGEM_42/43 insert
DNA	1 µl	3 µl	-	3 µl	1 µl	3 µl	-	3 µl
dH₂O	3 µl	1 µl	4 µl	1 µl	3 µl	1 µl	4 µl	1 µl
5xLD	1 µl	1 µl	1 µl	1 µl	1 µl	1 µl	1 µl	1 µl

(for Riboswitch iGEM_41/42 and _42/43, there were too small amounts of PCR product left -> control was not possible…)

Purification of DarR and Riboswitch inserts

Qiagen PCR purification kit
500 µl PB used for each reaction
2x elution in pre-warmed HPLC water (25 µl)
NanoDrop concentration measurements (for some samples, the concentration was measured twice, since first values for 1st and 2nd elution were a little strange… -> approximate average concentration can be used for further calculations)

1st elution

2nd elution

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