NanoBio: Archiving Your Plasmid in the Strain Collection

Archiving the New Part

• All new strains should be listed in our spreadsheet of strains and stored in the Molecular Foundry strain collection. It is also helpful to keep parts that you have made in your personal strain collection.
• It is recommended that you also add new plasmids to the plasmid collection.

Adding a Strain to the Strain Collection

1. Open Plasmid_Cell Stock Registry in Excel.
2. Give your plasmid a name.
   • Plasmids are names Lxxxx, where L is a letter and x is a number.
   • The letter indicates the function of the part. Currently
     • V=Vector Backbones
     • C=Binding Proteins
     • D=Delta (Knockout strains)
     • E=Reporter Proteins
     • G=Genetic Recombination Parts
     • H=Thermo Stability
     • I=Electron Transfer Proteins
     • J=Transporters
     • K=RBS's, translational parts
     • L=terminators, transcriptional modifiers
     • P=Promoters, binding sites
     • S=Structural proteins
     • T=tags
     • Z=Combination Parts
   • Use the next available number within a letter-sequence. For example, the next binding protein part after C5201 should be C5202.
3. Check that the next available MFe# has not already been used. To do so, use Ctrl-F (find) to see if that MFe# is already somewhere else in the spreadsheet.
4. Give the strain containing your plasmid a MFe#. Use the next available # as indicated at the top of the spreadsheet.
5. Update the next available #.
6. Insert a row (not just a series of cells) in the spreadsheet within the appropriate section, i.e. C for Binding proteins.
7. Fill in all the requested information into the spreadsheet
   • id#: the MFe number, e.g. MFe030
   • author: your initials, e.g. C.A-F
   • plasmids: the names of all plasmids in the Lxxxx format, e.g. T5015
   • brief 'part' description: VAAL-E4 peptide
   • vector backbone: the plasmid backbone, using the same name found in the vector section, i.e. pSB1AC3
   • Cell Line: the name of the cell strain, e.g. Top 10
   • More complete description:
   • size(bp): the size of just the part
   • biofusion or biobricks part?: if your part is in biofusion or biobricks format, please indicate here
   • VectorNTI file: the name of the vector NTI file, which should be the same as the plasmid name, i.e. T5015.
   • other notes:
   • Refs.: If this is described in a published paper, please insert that citation here.

Plasmid Collection

• Place 5 uL mini-prepped plasmid DNA in a labeled eppendorf tube in the appropriate plasmid collection freezer box in the -20 freezer in Rm 5204.

• For the strain collection, grow a bacterial culture either to mid-exponential phase or overnight. Mix equal volumes 50% glycerol and culture in an autoclaved 2 mL screw-top tube. Store at -80.

these are the blue labeled boxes on the Common Stocks shelf in the -80C in Rm 5204.