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Complete reference for all PrimerLab configuration parameters.

Configuration File Structure

input:
  sequence: "ATCGATCG..."  # or sequence_path
  sequence_path: /path/to/sequence.fasta

parameters:
  tm:
    min: 57.0
    opt: 60.0
    max: 63.0
  
  primer_size:
    min: 18
    opt: 20
    max: 25
  
  product_size:
    min: 100
    max: 300
  
  gc:
    min: 40.0
    max: 60.0

output:
  format: json  # json, csv, text
  directory: ./output

Parameter Reference

Input Parameters

ParameterTypeDescription
sequencestringRaw DNA sequence
sequence_pathpathPath to FASTA file

Tm (Melting Temperature)

ParameterDefaultRangeDescription
tm.min57.050-70Minimum Tm
tm.opt60.055-65Optimal Tm
tm.max63.058-72Maximum Tm

Primer Size

ParameterDefaultRangeDescription
primer_size.min1815-25Minimum length
primer_size.opt2018-22Optimal length
primer_size.max2520-30Maximum length

Product Size

ParameterDefaultDescription
product_size.min100Minimum amplicon
product_size.max300Maximum amplicon
Recommendations by workflow:
WorkflowMinMaxNotes
Standard PCR2001000Flexible
qPCR70150Short preferred
Long-range PCR100010000Specialized enzymes

GC Content

ParameterDefaultDescription
gc.min40.0Minimum GC%
gc.max60.0Maximum GC%

qPCR-Specific Parameters

parameters:
  probe:
    tm:
      min: 65.0
      opt: 68.0
      max: 72.0
    size:
      min: 20
      max: 30
ParameterDefaultDescription
probe.tm.opt68.0Probe Tm (5-10°C > primers)
probe.size.min20Minimum probe length
probe.size.max30Maximum probe length

Advanced Parameters

Salt Concentrations

parameters:
  salt:
    monovalent: 50.0   # mM (Na+, K+)
    divalent: 1.5      # mM (Mg2+)
    dntp: 0.2          # mM

Thermodynamic Thresholds

parameters:
  thermo:
    hairpin_dg_max: -2.0    # kcal/mol
    homodimer_dg_max: -5.0  # kcal/mol
    heterodimer_dg_max: -5.0

Output Configuration

output:
  format: json       # json, csv, text, html
  directory: ./output
  include_audit: true
  include_rationale: true

Example Configurations

Standard PCR

input:
  sequence_path: ./gene.fasta

parameters:
  tm:
    opt: 60.0
  product_size:
    min: 200
    max: 500

qPCR with TaqMan Probe

input:
  sequence_path: ./target.fasta

parameters:
  tm:
    opt: 60.0
  product_size:
    min: 70
    max: 150
  probe:
    tm:
      opt: 68.0

Long-Range PCR

input:
  sequence_path: ./region.fasta

parameters:
  tm:
    opt: 65.0
  product_size:
    min: 2000
    max: 10000
  primer_size:
    opt: 25

See Also