<< Worksheet Section 4.3 Analyzing Primer Pair Statistics >>

Action: Use Primer-3 to check Quality of your selected Primer pair.
Step 4.3 (Check your Primer Pair)

Analyzing Primer Pair Statistics

13. Please note the Primer Pair Statistic values in your document (cut & Paste).

Base pair position of first mtDNA mutation of working haplogroup :___________
Base pair position of last mtDNA mutation of working haplogroup :___________

Left primer "start" position plus primer length should be lower than base pair position for the first expected mutation.
Right primer "start" position minus primer length should be higher than the base pair position for the last expected mutation.

- Here is an example of the "Check-Primer" Output-table
       
PRIMER PICKING RESULTS FOR hap RSRS

No mispriming library specified
Using 1-based sequence positions
WARNING: Left primer is unacceptable: Tm too low; Right primer is unacceptable: High hairpin stability (thermod. approach)

OLIGO            start  len      tm     gc%  any_th  3'_th hairpin seq 
LEFT PRIMER      15972   20   56.91   50.00    0.00   0.00    0.00 TAACTCCACCATTAGCACCC
RIGHT PRIMER     16520   20   59.75   60.00    0.00   0.00   50.91 GGCCCTGAAGTAGGAACCAG
SEQUENCE SIZE: 16569
INCLUDED REGION SIZE: 16569

PRODUCT SIZE: 549, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00