Team:Saarland/Test

From 2014.igem.org

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<h1>Materials</h1>
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<iframe src="//player.vimeo.com/video/109005558" width="500" height="281" frameborder="0" webkitallowfullscreen mozallowfullscreen allowfullscreen></iframe> <p><a href="http://vimeo.com/109005558">Saarland University iGEM 2014 - Campus Tour</a> </p></html>
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<br>
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<!-- Inhaltsverzeichnis-->
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<h2>1. Chemicals </h2>
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<br>
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{|
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! Chemical
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! Manufacturer
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| 2 log DNA ladder
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| New England Biolabs
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| 2-Mercapto Ethanol
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| Merck
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| Acetic Acid 100%
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| zChL UdS
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| Agar Kobe I
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| Roth
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| Agarose
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| Peqlab, Biozym
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| Ampicillin 
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| Roth
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| APS (Ammoniumpersulfat)
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| zChL UdS
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| BactoTM Casamino-acids
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| BD
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| BactoTM Tryptone
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| BD
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| BactoTM Yeast-Extract
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| BD
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| Boric Acid
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| Merck
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| Bovine Serum Albumin (BSA)
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| New England Biolabs
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|-
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| Bromophenol Blue
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| Merck
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|-
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| Calcium Chloride
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| Calbiochem
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|-
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| Chloramphenicol
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| Fagran
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|-
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| D(+)-Glucose
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| Sigma Aldrich
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| ddH2O (MilliQ) (MILLIPORE 0,22 μm Membranfilter)
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| Biocel
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| DifcoTM Antibiotic Medium No. 3
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| BD
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| Dimethylsulfoxide (DMSO)
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| Sigma Aldrich
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| dNTP-Mix (10 mM)
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| Peqlab
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| EDTA
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| Roth
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| Ethanol, HPLC grade
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| Sigma Aldrich
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| Glutamate
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| Sigma Aldrich
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| Glycerol
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| zChL UdS
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| Hydrochloric Acid 37 % (HCl)
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| VWR
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| Immersion Oil
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| Leica
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| Isopropanol
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| Sigma Aldrich
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| LB-Medium (Miller)
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| Sigma Aldrich
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| Magnesium Chloride  Hexahydrate
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| VWR
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| Magnesium Sulfate
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| zChL UdS
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| Maleic Acid
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| Roth
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| Natriumchlorid
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| VWR
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| Natriumhydroxid
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| Grüssing
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| PEG-6000
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| Merck
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| peqGREEN
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| Peqlab
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| Potassium Chloride
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| Grüssing
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| Potassium Dihydrogen Orthophosphate
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| Grüssing
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| Potassium Hydrogen Phosphate
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| Grüssing
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| Potassium Sulfate
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| zChL UdS
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| Proline
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| Merck
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| Protein Standard
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| New England Biolabs
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| Rotiphorese Gel 30
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| Roth
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| SDS
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| Applichem/Roth
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| Succrose
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| Sigma Aldrich
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| Temed
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| Applichem
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| Tetracycline
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| Serva
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| Tris-Baze
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| Sigma Aldrich
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| Trizma Base
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| Sigma Aldrich
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| Xylose
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| Sigma Aldrich
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<h2> 2. Enzymes</h2>
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<br>
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{|
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!Enzyme
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!Manufacturer
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| Lysozyme
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| Serva
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| T4 Ligase
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| Thermo Scientific
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| Taq-Polymerase
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| New England Biolabs
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| Q5 High Fidelity Polymerase (Master Mix)
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| New England Biolabs
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| FastDigest <i>Mlu</i>I
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| Thermo Scientific
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| FastDigest <i>Sac</i>I
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| Thermo Scientific
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| FastDigest <i>Sph</i>I
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| Thermo Scientific
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| FastDigest <i>Eco</i>RI
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| Thermo Scientific
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| FastDigest <i>Xba</i>I
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| Thermo Scientific
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| FastDigest <i>Spe</i>I
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| Thermo Scientific
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| FastDigest <i>Pst</i>I
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| Thermo Scientific
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| Fast AP
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| Thermo Scientific
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<br>
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<h2> 3. oligonucleotides </h2>
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<br>
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{|
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!Abbreviation
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!5´-Sequence-3´
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| <b>Generation of <i>has2-gfp</i> fusion gene</b>
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| SOE Has for:
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| TGTACACGCGTAAATCAAGGAGGTG
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| SOE Has rev:
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| CTCGCCCTTGCTCACCATTACATCTAATACCATATCGTATTGTTGACC
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| SOE GFP for:
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| ATGGTATTAGATGTAATGGTGAGCAAGGGCGAG
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| SOE GFP rev:
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| TGTACGAGCTCTTACTTGTACAGCTCGTCCATGC
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|<b>Amplification of genomic DNA of <i>B. megaterium</i></b>
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| RBS UDP-GlcDH for
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| TGTACGAGCTCAAATCAAGGAGGTGAATATACAATGACTAATATCACAGTAGCGGGTACTGG
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| UDP-GlcDH rev:
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| TGTACGCATGCTTAGACATTAGCTTTAATTTTAGCTTTGTATCC
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| Glk for
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| TGTAC GAATTC ATGGATGACAAATGGTTAG
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| Glk rev:
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| TGTAC CTGCAG TTA TTATTCAAAATAAGAT
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| PgmB for
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| TGTAC GAATTC ATGAAAAAAGAAGAGCTAC
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| PgmB rev:
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|TGTAC ACTAGT TTA TTAATTTTTTGTATAC
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| GtaB for
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| TGTAC GAATTC ATGACGATAAAAAAGGCAG
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| GtaB rev:
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| TGTAC CTGCAG TTA TTAGCTGAAGTTGTTTGC
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| Pgi for
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| TGTAC TCTAGA ATGACACATGTTCGTTTTG
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| Pgi rev:
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| TGTAC CTGCAG TTA TTATTTCAAACGCTTTTC
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| GlmS for
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| TGTAC TCTAGA ATGTGCGGAATTGTAGG
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| GlmS rev:
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| TGTAC CTGCAG TTA TTATTCTACAGTAACTG
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| GlmM for
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| TGTAC GAATTC ATGGGTAAGTATTTTGG
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| GlmM rev:
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| TGTAC CTGCAG TTA TTACTCTAAGCCCATTTC
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| GcaD for
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| TGTAC GAATTC ATGTCAAAAAGATATGC
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| GcaD rev:
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| TGTAC CTGCAG TTA TTAGGATTTTTTATTAATATC
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|<b>Quick Change PCR for restriction site mutagenesis</b>
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| QC Has for:
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| GAATTTGTAGAAGATTGGTACTCTCAAGAATTTATGGGTAACCAG
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| QC has rev:
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| AAATTCTTGAGAGTACCAATCTTCTACAAATTCGTGTAATAAAGAG
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| QC UDP-GlcDH for:
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| TAATTATTAAAAACTCTGATGCTGGTAAATTATTTTTCACAACTG
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| QC UDP-GlcDH rev:
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| ACCAGCATCAGAGTTTTTAATAATTAATTCATCAAGCCCAGG
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| QC PgmB for:
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| TTGCAGCTGCTGATTGAGTTCTTCAG
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| QC PgmB rev:
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| CTGCAAGGGATCAGCCGAGTG
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| QC Pgi for:
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| AAATTCTTCTTTGTCATACTCCGTAGG
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| QC Pgi rev:
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| GAATTTGCACGCATTCAAAAAAGTGC
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| QC GlmS for:
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| AAATTCACTTGCTACGTGCACTTCTAC
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| QC GlmS rev:
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| GAATTTTCTTATAACATGCCGCTTTTATCTG
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| QC GlmM for:
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| TCGAGATCTGCAAATAAATGTGCTGC
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| QC GlmM for:
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| TCTCGATGCAGATATCTCTACAATGG
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|<b>Sequencing</b>
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| pSB1C3 check for:
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| TGCCACCTGACGTCTAAGAAACC
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| pSB1C3 check rev:
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| ATTACCGCCTTTGAGTGAGCTG
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<h2> 4. Media and Solutions </h2>
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!Solution
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!Ingredients
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!Quantity
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|10 x KPP Buffer
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|KH₂PO₄<br>
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K₂HPO₄<br>
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to 100 ml ddH2O
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autoclaved
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|2,71 g<br>
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12.5 g <br>
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</div>
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{{Team:Saarland/footer}}

Latest revision as of 12:39, 17 October 2014


Retrieved from "http://2014.igem.org/Team:Saarland/Test"