Team:Washington/Primers
From 2011.igem.org
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PCR Conditions and Primers of Registry Parts
Please fill out for every PCR reaction done on a registry part (both new and old).
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 |
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
EXAMPLE TABLE (DO NOT MODIFY)
Alphabetic | Numeric | Date | Unsortable |
---|---|---|---|
d | 20 | 2008-11-24 | This |
b | 8 | 2004-03-01 | column |
a | 6 | 1979-07-23 | cannot |
c | 4.2 | 1492-12-08 | be |
e | 0 | 1601-08-13 | sorted. |