Team:Washington/Primers

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(PCR Conditions and Primers of Registry Parts)
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| S || BBa_XYZ001 || M || Synthesized Gene || EX_Decarb_F || RedDecarb_SP_2 || 60 || 30 || 0 || Phusion || GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCCGCAGCTGGAA || CCAATGCATTGGTTCTGCAGCGGCCGCTACTAGTATCAGTGGTGGTGGTG || Original reverse primer was missing extra nucleotides at end, so amplification worked, cloning didn't
| S || BBa_XYZ001 || M || Synthesized Gene || EX_Decarb_F || RedDecarb_SP_2 || 60 || 30 || 0 || Phusion || GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCCGCAGCTGGAA || CCAATGCATTGGTTCTGCAGCGGCCGCTACTAGTATCAGTGGTGGTGGTG || Original reverse primer was missing extra nucleotides at end, so amplification worked, cloning didn't
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| S || ### ||M || Gene from Collaborator || KumaCloning-F || KumaCloning-R || 66 || 60 || 0 || Phusion || GTTTCTTCGAATTCGCGGCCG || GTTTCTTTCCTGCAGCGGCCGG || These primers amplify the Kumamolisin gene, adding standard BioBrick ends for cloning.
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Revision as of 20:59, 14 September 2011

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Contents

PCR Conditions and Primers of Registry Parts

Please fill out for every PCR reaction done on a registry part (both new and old).

PCR Conditions and Primers of Registry Parts
Amplification Success
(S, M, N)
Template Part # Template Form
(M, C, P)
Template Source Forward Primer Name Reverse Primer Name Annealing Temperature
(Celsius)
Extension Time
(seconds)
Percent DMSO Polymerase Used Forward Primer
(5'->3')
Reverse Primer
(5'->3')
Additional Notes
S BBa_XYZ001 M Synthesized Gene EX_Decarb_F RedDecarb_SP_2 60 30 0 Phusion GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCCGCAGCTGGAA CCAATGCATTGGTTCTGCAGCGGCCGCTACTAGTATCAGTGGTGGTGGTG Original reverse primer was missing extra nucleotides at end, so amplification worked, cloning didn't
S ### M Gene from Collaborator KumaCloning-F KumaCloning-R 66 60 0 Phusion GTTTCTTCGAATTCGCGGCCG GTTTCTTTCCTGCAGCGGCCGG These primers amplify the Kumamolisin gene, adding standard BioBrick ends for cloning.
Fill Fill Fill Fill Fill Fill Fill Fill Fill Fill Fill Fill Fill

LEGEND

  • Amplification Success (S, M, N): Was the amplification successful.
    • S = Single Band at Desired Size
    • M = Multiple Bands, but one at desired length
    • N = No band at desired length
  • Template Part #: The Registry Number of the Part being amplified from
  • Template Form: The source form of the template DNA
    • Miniprep (M)
    • Colony (C)
    • PCR Product (P)
  • Template Source: Where the template is originally from, such as 2011 Parts Kits, Synthesized Gene, Genomic, etc
  • Forward Primer (5'->3'): Full sequence of the Forward primer used
  • Reverse Primer (5'->3'): Full sequence of the Reverse primer used
  • Annealing Temperature (Celsius): The annealing temperature used in the PCR reaction
  • Extension Time (seconds): The extension time used in the PCR reaction
  • Percent DMSO: Was DMSO added, if not 0, if so at what %
  • Polymerase Used: Which polymerase enzyme was used (e.g. Phusion, Taq, Vent, Pfu, etc)
  • Primer Name: The name of the primer ordered
  • Additional Notes: Any additional relevant information

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