From 2012.igem.org
(Difference between revisions)
Revision as of 11:14, 23 September 2012
Materials
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
|
Enzymes
| No.
| Enzyme
| Supplier
|
| 1
| Antarctic Phosphatase
| NEB
|
| 2
| BsaI
| NEB
|
| 3
| EcoRI-HF
| NEB
|
| 4
| Phusion® High-Fidelity DNA Polymerase
| NEB
|
| 5
| PstI-HF
| NEB
|
| 6
| SpeI-HF
| NEB
|
| 7
| T4 DNA Ligase
| NEB
|
| 8
| Taq DNA Polymerase with ThermoPol Buffer
| NEB
|
| 9
| XbaI
| NEB
|
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
|
Kits
| No.
| Kit
| Supplier
|
| 1
| PureYield™ Plasmid Miniprep System
| Promega
|
| 2
| PureYield™ Plasmid Midiprep System
| Promega
|
| 3
| Wizard® SV Gel and PCR Clean-Up System
| Promega
|
Consumabels
| No.
| Article
| Supplier
|
| 1
| epT.I.P.S.® Box pipette tips 0.1–20 µL
| Eppendorf
|
| 2
| epT.I.P.S.® Box pipette tips 10–200 µL
| Eppendorf
|
| 3
| epT.I.P.S.® Box pipette tips 100–1000 µL
| Eppendorf
|
| 4
| epT.I.P.S.® Bulk pipette tips 0.1–20 µL
| Eppendorf
|
| 5
| epT.I.P.S.® Bulk pipette tips 10–200 µL
| Eppendorf
|
| 6
| epT.I.P.S.® Bulk pipette tips 100–1000 µL
| Eppendorf
|
| 7
| PCR Tubes 0.2 mL
| Eppendorf
|
| 8
| PCR Tube Strips 0.2 mL
| Eppendorf
|
| 9
| Safe-Lock Tubes (1.5 mL)
| Eppendorf
|
| 10
| Safe-Lock Tubes (2 mL)
| Eppendorf
|
| 11
| Rotilabo®-syringe filters
| Carl Roth GmbH
|
| 12
| NORM-Ject Syringes
| Henke/Sass/Wolf
|
| 13
| Multiply®-PCR-tubes 0.2 mL
| Sarstedt
|
| 14
| VWR® Pipet Tips (Standard Tips, 0.1–20 µL)
| VWR
|
| 15
| VWR® Pipet Tips (Standard Tips, 10–200 µL)
| VWR
|
| 16
| VWR® Pipet Tips (Standard Tips, 100-1000 µL)
| VWR
|
| 17
| VWR® Powder-Free Nitrile Examination Gloves
| VWR
|
| 18
| Eco-friendly centrifuge tubes (15 mL, unsterile)
| Carl Roth GmbH
|
| 19
| 50 mL Centrifuge tubes, sterile
| Greiner bio-one
|
| 20
| Sekuroka®-disposal bags
| Carl Roth GmbH
|
| 21
| Rotiprotect®-nitrile gloves, unpowdered
| Carl Roth GmbH
|
| 22
| UV-cuvettes PLASTIBRAND® (Micro z = 8,5 mm)
| Brand GmbH & Co. KG
|
| 23
| Parafilm® M
| Pechiney Plastic Packaging
|
Equipment
| No.
| Equipment
| Supplier
|
| 1
| Mastercycler personal (Thermocycler)
| Eppendorf
|
| 2
| Heraeus Pico Microcentrifuge
| Thermo Scientific
|
| 3
| Multifuge® 1S-R
| Thermo Scientific
|
| 4
| VX-150 Autoclave
| Systec
|
| 5
| Uni-Drybath Thermoshaker
| Universal Labortechnik
|
| 6
| Vortex Genie 2
| Scientific Industries
|
| 7
| Universal Centrifuge
| Sigma Laborzentrifugen
|
| 8
| Dark Hood DH-40 with control panel and display
| Biostep
|
| 9
| BioMate 3S Spectrophotometer
| Thermo Scientific
|
| 10
| GENESYS 10S UV-Vis
| Thermo Scientific
|
| 11
| Heraeus LaminAir HA 2448 GS
| Heraeus
|
| 12
| Signature™ Ergonomic High Performance Single-Channel Variable Volume Pipettors 10 µL
| VWR
|
| 13
| Signature™ Ergonomic High Performance Single-Channel Variable Volume Pipettors 100 µL
| VWR
|
| 14
| Signature™ Ergonomic High Performance Single-Channel Variable Volume Pipettors 1000 µL
| VWR
|
| 15
| Research® plus, adjustable pipette 0.5 - 10 µL
| Eppendorf
|
| 16
| Research® plus, adjustable pipette 10 - 100 µL
| Eppendorf
|
| 17
| Research® plus, adjustable pipette 100 – 1000 µL
| Eppendorf
|
| 18
| Research® plus, adjustable pipette 10 - 200 µL
| Eppendorf
|
| 19
| Research® plus, adjustable pipette 0.5 - 20 µL
| Eppendorf
|
| 20
| Reference, adjustable pipette 0.5 – 10 µL
| Eppendorf
|
| 21
| M-Prove Balances
| Sartorius
|
| 22
| M-Power Balances
| Sartorius
|
| 23
| Owl Series EasyCast™ Horizontal Mini-Gel Systems
| VWR
|
| 24
| Owl Series Horizontal Gel Systems
| VWR
|
| 25
| Owl Series Dual Gel Vertical Electrophoresis System
| VWR
|
| 26
| Power Supplies 250V
| VWR
|
| 27
| Incubation/Inactivation Water Bath 1008
| Gesellschaft für Labortechnik
|
| 28
| Lab-Shaker LSR-V
| Adolf Kühner AG
|
| 29
| MR Hei-Mix S
| Heidolph
|
| 30
| FE20 – FiveEasy™ pH
| Mettler Toledo
|
"
