Team:Goettingen/NoteBook w7

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Revision as of 14:14, 16 September 2013

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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