Team:TU Darmstadt/Project/Metabolism/Materials

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Protocols Metabolism

Contents

Chemicals

No. Name Firma Order-ID
1 Terephthalat Sigma-Aldrich 185361-500G
2 Acetic acid Roth 6755.1
3 Methanol Roth CP43.1
4 Coomassie Brilliant Blue R-250 Roth 9598.1
5 Trypton Roth 8952.1
6 Ethylene glycol Roth 9516.1
7 3,4-Dihydroxybenzoic acid Sigma-Aldrich 37580-25G-F
8 Catechol Sigma-Aldrich C9510-100G
9 Isopropyl β-D-1-thiogalactopyranoside Roth CN08.1
10 Tetramethylethylenediamine Roth 2367.3
11 Ammonium persulfate Roth 9592.3
12 Bromophenol blue Roth A512.1
13 Glucose Roth X997.1
14 Calcium chloride Roth CN92.1
15 Dithiothreitol Roth 6908.1
16 Sodium carbonate Roth 8563.1
17 Phenylmethanesulfonylfluoride Roth 6367.1
18 NADH-Disodium Roth AE12.1
19 Nicotinamide adenine dinucleotide Roth AE11.1
20 orthophosphoric acid 85% Roth 9079.1
21 Trichloroacetic acid Roth 8789.2
22 Quick-Load® 100 bp DNA Ladder NEB Biolabs N0467S
23 Phenol AppliChem A4458.0250
24 Dimethyl sulfoxide AppliChem A1584.0100
25 Methylene blue AppliChem A4084.0025
26 Magnesium sulfate AppliChem A6287.0250
27 Cetrimonium bromide AppliChem A0805.0100
28 Tetramethylethylenediamine AppliChem A1148.0025
29 Urea AppliChem A5470.0500
30 Tween20 AppliChem A1389.0500
31 Protein Marker III (6,5-200) AppliChem A4402.0001
32 Thiamine hydrochloride AppliChem A09955.0050
33 Triton X-100 AppliChem A1388.0500
34 DNA-Dye NonTox AppliChem A9555.1000
35 Agarose Roth 3810.2
36 Ethanol 96 % Roth T171.4
37 Ethylenediaminetetraacetic acid Roth X986.2
38 Ethanol Roth P076.1
39 Tetracycline hydrochloride Roth 237.1
40 D-Desthiobiotin 10x Buffer E iba 2-1000-025
41 2-(4-Hydroxyphenylazo)benzoic acid Sigma-Aldrich H5126-5G
42 10 mM dNTP-Mix Roth L785.2
43 Hydrochloric acid Roth 9277.1
44 Acrylamid - Bis (37,5:1) Roth 3029.1
45 Ammonium acetate Roth 7869.2
46 Agar-Agar Roth 5210.3
47 Chloroform Roth 4423.1
48 Glycine Roth 3790.2
49 Dipotassium phosphate Roth T875.1
50 Sodium chloride Roth 9265.1
51 Yeast extract Roth 2363.2
52 Monopotassium phosphate Roth P018.1
53 Glycerine Roth 3783.1
54 Sodium hydroxide Roth P031.1
55 Tris-Base Roth AE15.2
56 Isopropyl alcohol Roth CP41.3
57 Isoamyl alcohol Roth T870.1
58 Acetonitrile Roth 7330.2
59 Acetone Roth 5025.5
60 Chloramphenicol Roth 3886.2
61 Ampicillin Roth K029.1
62 Kanamycin Roth T832.1
63 Sodium dodecyl sulfate Roth 5136.1

Primer

No. Name 5´-->3´
1 tphA2-l-F TCATGCTTGCATCTCCTGTC
2 tphA2-l-Prefix GAATTCGCGGCCGCTTCTAGATGCAAGAATCCATCATCCAGTGGC
3 tphA1-Suffix_R CTGCAGCGGCCGCTACTAGTACTAATGGTTGCCAGTCGGGTCTG
4 tphA3-Suffix_R CTGCAGCGGCCGCTACTAGTATCATAGCGGCAATGACATCAGCGTG
5 tphA3-Prefix_F GAATTCGCGGCCGCTTCTAGATGATCCATGAAATTCAAATCGCGG
6 tphA1-l-R CTAATGGTTGCCAGTCGGGT
7 tphA1-l-PstI(99)-F CATGGTCTCCTCCAGGCCGGCATCGAGCT
8 tphA1-l-Prefix GAATTCGCGGCCGCTTCTAGATGAACCACCAGATCCATATCCACG
9 tphA3-l-F TCATAGCGGCAATGACATCA
10 tphA3-l-Prefix GAATTCGCGGCCGCTTCTAGAGATGATCCATGAAATTCAAATCGC
11 Prefix GAATTCGCGGCCGCTTCTAGAG
12 tphB-l-F TCAGACCGGTTGGGCTCCGA
13 tphB-l-Prefix GAATTCGCGGCCGCTTCTAGAGATGACAATAGTGCACCGTAGATT
14 tphA2-l-R ATGCAAGAATCCATCATCCAG
15 tphA1-l-R ATGAACCACCAGATCCATATC
16 tphA1-l-PstI(99)-R CATGGTCTCCTGGAGGGCTGCATCCAGTAC
17 tphA3-l-R ATGATCCATGAAATTCAAAT
18 Suffix CTGCAGCGGCCGCTACTAGTA
19 tphB-l-Suffix-R CTGCAGCGGCCGCTACTAGTATCAGACCGGTTGGGCTCCGAGTA
20 EcoRIGFxa-tphA1 TTCTGGAATTCGGGTATTGAAGGAAGAATGAACCACCAGATCCATAT
21 EcoRIGFxa-tphA2 TTCTGGAATTCGGGTATTGAAGGAAGAATGCAAGAATCCATCATCCA
22 EcoRIGFxa-tphA3 TTCTGGAATTCGGGTATTGAAGGAAGAATGATCCATGAAATTCAAAT
23 EcoRIGFxa-tphB TTCTGGAATTCGGGTATTGAAGGAAGAATGACAATAGTGCACCGTAG
24 EcoRIGFxa-aroY TTCTGGAATTCGGGTATTGAAGGAAGAATGACAGCGCCTATCCAG
25 RBS-tphA1 TTCTGGAATTCGCGGCCGCTTCTAGAGAAAGACAGGACCTACTAGATGAACCACCAGATCCATAT
26 RBS-tphA2 TTCTGGAATTCGCGGCCGCTTCTAGAGAAAGACAGGACCTACTAGATGCAAGAATCCATCATCCA
27 RBS-tphA3 TTCTGGAATTCGCGGCCGCTTCTAGAGAAAGACAGGACCTACTAGATGATCCATGAAATTCAAAT
28 RBS-tphB TTCTGGAATTCGCGGCCGCTTCTAGAGAAAGACAGGACCTACTAGATGACAATAGTGCACCGTAG
29 RBS-aroY TTCTGGAATTCGCGGCCGCTTCTAGAGAAAGACAGGACCTACTAGATGACAGCGCCTATCCAG
30 VF2 TGCCACCTGACGTCTAAGAA
31 VR ATTACCGCCTTTGAGTGAGC
32 pPR-IBA2 rev TAGTTATTGCTCAGCGGTGG
33 pPR-IBA2 for TAATACGACTCACTATAGGG

Plasmids

No. Name Sequence
1 pSB1C3 [http://partsregistry.org/Part:pSB1C3 Get]
2 pSB1AK3 [http://partsregistry.org/Part:pSB1AK3 Get]
3 BBa_J61002 [http://partsregistry.org/Part:BBa_J61002 Get]
4 pSB1A2 [http://partsregistry.org/Part:pSB1A2 Get]
5 pPR-IBA2 [http://www.iba-lifesciences.com/isotope/2/2-1391-000-Sequence-pPR-IBA2.txt Get]

Biobricks from the registry

No. Name Platte Well Resistence Plasmid backbone
1 BBa_K316003 4 15C C pSB1C3
2 BBa_J23100 1 18C A BBa_J61002
3 BBa_B0015 1 23L A und K pSB1AK3
4 BBa_J61101 1 5L A pSB1A2


Bacteria

Species Strain Genotype DSM-No.
Escherichia coli DH5α F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(rK- mK+), λ– 6897
Escherichia coli MG1655 F- λ- ilvG- rfb-50 rph-1 18039
Escherichia coli BL21(DE3)pLysS F- ompT gal dcm lon hsdSB(rB- mB-) λ(DE3) pLysS(cmR) none
Comamonas testosteroni Kf-1 - 14576
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