Team:Bielefeld-Germany/Protocols/Materials

From 2012.igem.org

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==Primers==
==Primers==
This is a list of primers we have used.
This is a list of primers we have used.
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 +
{| class="wikitable"
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|-
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! primer name !! lenght !! sequence !!
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|-
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| pSB1C3-5aox1-f || 60 || CGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGAGATCTAACATCCAAAGACG
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|-
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| pSB1C3-5aox1-r || 30 || GGTGGCGGCGGGCGTTTCGAATAATTAGTT
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|-
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| 5aox1-mfalpha1-f || 68 || AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC
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|-
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| 5aox1-mfalpha1-r || 20 || AGCTTCAGCCTCTCTTTTCT
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|-
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| mfalpha1-aarI-taox1-f || 80 || GTATCTCTCGAGAAAAGAGAGGCTGAAGCTACACGCAGGTGGTATGTATCACCTGCGTGTCTTGCTAGATTCTAATCAAG
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|-
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| mfalpha1-aarI-taox1-r || 20 || TAAGCTTGCACAAACGAACT
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|-
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| taox1-phis4-f || 60 || GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTATCATGCCATGGACAAGATTC
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|-
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| taox1-phis4-r || 20 || GGCCGCTCGAGTATTCAGAA
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|-
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| phis4-kozak-his4-f || 72 || AATAGTTTACAAAATTTTTTTTCTGAATACTCGAGCGGCCCCCGCCGCCACCATGACATTTCCCTTGCTACC
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|-
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| phis4-kozak-his4-r || 30 || TTATTATTTCTCCATACGAACCTTAACAGC
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|-
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| his4-3aox1-f || 60 || TCACCGCAATGCTGTTAAGGTTCGTATGGAGAAATAATAACGAGTATCTATGATTGGAAG
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|-
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| his4-3aox1-r || 20 || AAAACAAGATAGTGCCCCTC
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|-
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| 3aox1-pSB1C3-f || 60 || AGTCTGATCCTCATCAACTTGAGGGGCACTATCTTGTTTTTACTAGTAGCGGCCGCTGCA
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|-
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| 3aox1-pSB1C3-r || 20 || CTCTAGAAGCGGCCGCGAAT
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|-
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| taox-his4-f || 61 || GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC
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|-
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| taox-his4-r || 27 || CTCGGATCTATCGAATCTAAATGTAAG
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|-
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| his4-3aox1-f02 || 60 || TTATTTAGAGATTTTAACTTACATTTAGATTCGATAGATCCGAGTATCTATGATTGGAAG
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|-
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| his4_gi537483_f || 46 || ACGTgaattcgcggccgcttctagagAGATCTCCTGATGACTGACT
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|-
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| his4_gi537483_r || 41 || ctgcagcggccgctactagtaGATCTATCGAATCTAAATGT
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|-
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|}

Revision as of 17:30, 23 August 2012

Contents

Materials

This is where we are going to list all our materials, devices and equipment that we have used.


Media, buffer and other solutions

Ampicillin stock solution

  • Solubilize 100 mg mL-1 Ampicillin
  • Store at -20 °C


Chloramphenicol stock solution

  • Solubilize 20 mg mL-1 Chloramphenicol in 100 % Ethanol
  • Store at -20 °C


TAE buffer

For 1 L of 50 x TAE buffer you need:

  • 242.48 g Tris
  • 41.02 g Sodiumacetate
  • 18.612 g EDTA
  • Adjust pH to 7.8 with acetic acid
  • Solve in dH2O

10 mL of the stock is diluted in 1 L dH2O for the gel electrophoresis (0.5 x TAE buffer).


DNA loading buffer

  • 50 % (v/v) glycerol
  • 1 mM EDTA
  • 0.1 % (w/v) bromphenol blue
  • Solve in ddH2O


LB media

For 1 L of LB media:

  • 10 g Trypton
  • 5 g Yeast extract
  • 10 g NaCl
  • 12 g Agar-Agar (for plates)
  • Adjust pH to 7.4


YPD media

For 1 L of YPD media:

  • 20 g Peptone
  • 10 g Yeast extract
  • 20 g Dextrose (add 50 mL sterile stock solution (40% dextrose))
  • Adjust pH to 6.5


Primers

This is a list of primers we have used.

primer name lenght sequence
pSB1C3-5aox1-f 60 CGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGAGATCTAACATCCAAAGACG
pSB1C3-5aox1-r 30 GGTGGCGGCGGGCGTTTCGAATAATTAGTT
5aox1-mfalpha1-f 68 AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC
5aox1-mfalpha1-r 20 AGCTTCAGCCTCTCTTTTCT
mfalpha1-aarI-taox1-f 80 GTATCTCTCGAGAAAAGAGAGGCTGAAGCTACACGCAGGTGGTATGTATCACCTGCGTGTCTTGCTAGATTCTAATCAAG
mfalpha1-aarI-taox1-r 20 TAAGCTTGCACAAACGAACT
taox1-phis4-f 60 GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTATCATGCCATGGACAAGATTC
taox1-phis4-r 20 GGCCGCTCGAGTATTCAGAA
phis4-kozak-his4-f 72 AATAGTTTACAAAATTTTTTTTCTGAATACTCGAGCGGCCCCCGCCGCCACCATGACATTTCCCTTGCTACC
phis4-kozak-his4-r 30 TTATTATTTCTCCATACGAACCTTAACAGC
his4-3aox1-f 60 TCACCGCAATGCTGTTAAGGTTCGTATGGAGAAATAATAACGAGTATCTATGATTGGAAG
his4-3aox1-r 20 AAAACAAGATAGTGCCCCTC
3aox1-pSB1C3-f 60 AGTCTGATCCTCATCAACTTGAGGGGCACTATCTTGTTTTTACTAGTAGCGGCCGCTGCA
3aox1-pSB1C3-r 20 CTCTAGAAGCGGCCGCGAAT
taox-his4-f 61 GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC
taox-his4-r 27 CTCGGATCTATCGAATCTAAATGTAAG
his4-3aox1-f02 60 TTATTTAGAGATTTTAACTTACATTTAGATTCGATAGATCCGAGTATCTATGATTGGAAG
his4_gi537483_f 46 ACGTgaattcgcggccgcttctagagAGATCTCCTGATGACTGACT
his4_gi537483_r 41 ctgcagcggccgctactagtaGATCTATCGAATCTAAATGT
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