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From 2012.igem.org

Contents 1 Materials 1.1 Media, buffer and other solutions 1.1.1 Ampicillin stock solution 1.1.2 Chloramphenicol stock solution 1.1.3 TAE buffer 1.1.4 DNA loading buffer 1.1.5 LB media 1.1.6 YPD media 1.2 Primers

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	length	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