Team:Saarland/Test
From 2014.igem.org
(Difference between revisions)
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<br> | <br> | ||
- | <h2> 3. | + | <h2> 3. oligonucleotides </h2> |
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{| | {| | ||
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| SOE GFP rev: | | SOE GFP rev: | ||
| TGTACGAGCTCTTACTTGTACAGCTCGTCCATGC | | TGTACGAGCTCTTACTTGTACAGCTCGTCCATGC | ||
- | |||
|- | |- | ||
+ | |<b>Amplification of genomic DNA of <i>B. megaterium</i></b> | ||
+ | | | ||
+ | |- | ||
+ | | RBS UDP-GlcDH for | ||
+ | | TGTACGAGCTCAAATCAAGGAGGTGAATATACAATGACTAATATCACAGTAGCGGGTACTGG | ||
+ | |- | ||
+ | | UDP-GlcDH rev: | ||
+ | | TGTACGCATGCTTAGACATTAGCTTTAATTTTAGCTTTGTATCC | ||
+ | |- | ||
+ | | Glk for | ||
+ | | TGTAC GAATTC ATGGATGACAAATGGTTAG | ||
+ | |- | ||
+ | | Glk rev: | ||
+ | | TGTAC CTGCAG TTA TTATTCAAAATAAGAT | ||
+ | |- | ||
+ | | PgmB for | ||
+ | | TGTAC GAATTC ATGAAAAAAGAAGAGCTAC | ||
+ | |- | ||
+ | | PgmB rev: | ||
+ | |TGTAC ACTAGT TTA TTAATTTTTTGTATAC | ||
+ | |- | ||
+ | | GtaB for | ||
+ | | TGTAC GAATTC ATGACGATAAAAAAGGCAG | ||
+ | |- | ||
+ | | GtaB rev: | ||
+ | | TGTAC CTGCAG TTA TTAGCTGAAGTTGTTTGC | ||
+ | |- | ||
+ | | Pgi for | ||
+ | | TGTAC TCTAGA ATGACACATGTTCGTTTTG | ||
+ | |- | ||
+ | | Pgi rev: | ||
+ | | TGTAC CTGCAG TTA TTATTTCAAACGCTTTTC | ||
+ | |- | ||
+ | | GlmS for | ||
+ | | TGTAC TCTAGA ATGTGCGGAATTGTAGG | ||
+ | |- | ||
+ | | GlmS rev: | ||
+ | | TGTAC CTGCAG TTA TTATTCTACAGTAACTG | ||
+ | |- | ||
+ | | GlmM for | ||
+ | | TGTAC GAATTC ATGGGTAAGTATTTTGG | ||
+ | |- | ||
+ | | GlmM rev: | ||
+ | | TGTAC CTGCAG TTA TTACTCTAAGCCCATTTC | ||
+ | |- | ||
+ | | GcaD for | ||
+ | | TGTAC GAATTC ATGTCAAAAAGATATGC | ||
+ | |- | ||
+ | | GcaD rev: | ||
+ | | TGTAC CTGCAG TTA TTAGGATTTTTTATTAATATC | ||
+ | |- | ||
+ | |||
Revision as of 12:34, 8 October 2014
Materials
1. Chemicals
Chemical | Manufacturer |
---|---|
2 log DNA ladder | New England Biolabs |
2-Mercapto Ethanol | Merck |
Acetic Acid 100% | zChL UdS |
Agar Kobe I | Roth |
Agarose | Peqlab, Biozym |
Ampicillin | Roth |
APS (Ammoniumpersulfat) | zChL UdS |
BactoTM Casamino-acids | BD |
BactoTM Tryptone | BD |
BactoTM Yeast-Extract | BD |
Boric Acid | Merck |
Bovine Serum Albumin (BSA) | New England Biolabs |
Bromophenol Blue | Merck |
Calcium Chloride | Calbiochem |
Chloramphenicol | Fagran |
D(+)-Glucose | Sigma Aldrich |
ddH2O (MilliQ) (MILLIPORE 0,22 μm Membranfilter) | Biocel |
DifcoTM Antibiotic Medium No. 3 | BD |
Dimethylsulfoxide (DMSO) | Sigma Aldrich |
dNTP-Mix (10 mM) | Peqlab |
EDTA | Roth |
Ethanol, HPLC grade | Sigma Aldrich |
Glutamate | Sigma Aldrich |
Glycerol | zChL UdS |
Hydrochloric Acid 37 % (HCl) | VWR |
Immersion Oil | Leica |
Isopropanol | Sigma Aldrich |
LB-Medium (Miller) | Sigma Aldrich |
Magnesium Chloride Hexahydrate | VWR |
Magnesium Sulfate | zChL UdS |
Maleic Acid | Roth |
Natriumchlorid | VWR |
Natriumhydroxid | Grüssing |
PEG-6000 | Merck |
peqGREEN | Peqlab |
Potassium Chloride | Grüssing |
Potassium Dihydrogen Orthophosphate | Grüssing |
Potassium Hydrogen Phosphate | Grüssing |
Potassium Sulfate | zChL UdS |
Proline | Merck |
Protein Standard | New England Biolabs |
Rotiphorese Gel 30 | Roth |
SDS | Applichem/Roth |
Succrose | Sigma Aldrich |
Temed | Applichem |
Tetracycline | Serva |
Tris-Baze | Sigma Aldrich |
Trizma Base | Sigma Aldrich |
Xylose | Sigma Aldrich |
2. Enzymes
Enzyme | Manufacturer |
---|---|
Lysozyme | Serva |
T4 Ligase | Thermo Scientific |
Taq-Polymerase | New England Biolabs |
Q5 High Fidelity Polymerase (Master Mix) | New England Biolabs |
FastDigest MluI | Thermo Scientific |
FastDigest SacI | Thermo Scientific |
FastDigest SphI | Thermo Scientific |
FastDigest EcoRI | Thermo Scientific |
FastDigest XbaI | Thermo Scientific |
FastDigest SpeI | Thermo Scientific |
FastDigest PstI | Thermo Scientific |
Fast AP | Thermo Scientific |
3. oligonucleotides
Abbreviation | 5´-Sequence-3´ |
---|---|
Generation of has2-gfp fusion gene | |
SOE Has for: | TGTACACGCGTAAATCAAGGAGGTG |
SOE Has rev: | CTCGCCCTTGCTCACCATTACATCTAATACCATATCGTATTGTTGACC |
SOE GFP for: | ATGGTATTAGATGTAATGGTGAGCAAGGGCGAG |
SOE GFP rev: | TGTACGAGCTCTTACTTGTACAGCTCGTCCATGC |
Amplification of genomic DNA of B. megaterium | |
RBS UDP-GlcDH for | TGTACGAGCTCAAATCAAGGAGGTGAATATACAATGACTAATATCACAGTAGCGGGTACTGG |
UDP-GlcDH rev: | TGTACGCATGCTTAGACATTAGCTTTAATTTTAGCTTTGTATCC |
Glk for | TGTAC GAATTC ATGGATGACAAATGGTTAG |
Glk rev: | TGTAC CTGCAG TTA TTATTCAAAATAAGAT |
PgmB for | TGTAC GAATTC ATGAAAAAAGAAGAGCTAC |
PgmB rev: | TGTAC ACTAGT TTA TTAATTTTTTGTATAC |
GtaB for | TGTAC GAATTC ATGACGATAAAAAAGGCAG |
GtaB rev: | TGTAC CTGCAG TTA TTAGCTGAAGTTGTTTGC |
Pgi for | TGTAC TCTAGA ATGACACATGTTCGTTTTG |
Pgi rev: | TGTAC CTGCAG TTA TTATTTCAAACGCTTTTC |
GlmS for | TGTAC TCTAGA ATGTGCGGAATTGTAGG |
GlmS rev: | TGTAC CTGCAG TTA TTATTCTACAGTAACTG |
GlmM for | TGTAC GAATTC ATGGGTAAGTATTTTGG |
GlmM rev: | TGTAC CTGCAG TTA TTACTCTAAGCCCATTTC |
GcaD for | TGTAC GAATTC ATGTCAAAAAGATATGC |
GcaD rev: | TGTAC CTGCAG TTA TTAGGATTTTTTATTAATATC |