Edinburgh/Lab

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Revision as of 15:48, 2 August 2007

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Biobricks Revived

This is a list of the biobricks we have revived from the registry

Lab Diary

Thu 12th July

• IPTG induced GFP revived ([http://partsregistry.org/Part:BBa_J04430 BBa_J04430])

Test biobrick revival process

Fri 13th July

• Alfalfa Seeds Planted (in Agar)

Growing alfalfa plants, to get leaves from which to extract DNA for the caffeic acid-3-o-methyltransferase gene (part of the vanillin synthesis pathway)

• Biobricks [http://partsregistry.org/Part:BBa_E0241 BBa_E0241] and [http://partsregistry.org/Part:BBa_J45700 BBa_J45700] revived, using modified protocal from open wetware site

Mon 16th July

• Begin DNA extraction from Bamboo

Test DNA extraction process from plants

• Prepared mini preps for GFP and mint genes

Tue 17th July

• Alfalfa Transferred to compost

• DNA containing GFP and mint genes extracted from E.Coli

Wed 18th July

• Double digest GFP and mint mini preps, using restriction enzymes SpeI and EcoRI. These were then run on a polyacramide gel
• Biobrick ECFP [http://partsregistry.org/Part:BBa_E0422 BBa_E0422] revived and transformed into E.coli JM109 competent cells

Thu 19th July

• Repeat of double digest (using SpeI & EcoRI) of GFP and mint mini preps with increased concentration of DNA
• Replate colonies of ECFP, 4 overnight cultures prepared for making DNA mini preps
• designed primers for COMT gene (part of vanillin biosynthesis pathway)

Fri 20th July

• Prepared GFP and winter green biobricks for sequencing
• Revived Pseudamonas fluorescens from freeze dried culture (P. fluorescens contains two genes, ech and fcs, which are part of the synthetic vanillin biosynthesis pathway)
• Made 4 mini preps of ECFP
• Designed primers for ech and fcs genes
• Ordered Saccharothrix espeanensis from DSMZ culture collection, which contains Sam5 and Sam8 genes required for the synthetic vanillin biosynthesis pathway

Mon 23rd July

• Ran gels for ECFP mini preps

Restriction digested using XbaI and PstI

• Alfalfa growing well - now have enough leaf material from which to extract DNA

Tue 24th July

• Annealed dif F & R oligoes, restriction digested with SacI & SpeI, ligated with BioBrick vector Edinbrick 1

Wed 25th July

• Transformed dif F & R DNA into E.coli, incubated at 37C overnight
• designed primers for Sam5 and Sam8 genes (part of vanillin biosynthesis pathway)

Thu 26th July

• Prepared glycerol stock of Pseudomonas fluorescens
• Prepared miniprep of GFP and mint biobricks
• Streaked Pseudomonas fluorescens onto a fresh LB plate
• Revived [http://partsregistry.org/Part:BBa_I0500 BBa_I0500] and [http://partsregistry.org/Part:BBa_Q04510 BBa_Q04510]

Fri 27th July

• Continued miniprep of GFP and mint biobricks

Restriction digest using SpeI & EccRI restriction enzymes

• PCR of Pseudomonas fluorescens genes