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Lab Manual
Sameer Khanal
available at: https://www.slideshare.net/SameerKhanal1
Linkage Mapping using MapMaker
Software download site:
http://research.franklin.uga.edu/devoslab/scripts-used-genetic-mapping
Latest version: MapMaker/EXP 3.0b January 1993
Destination: closest to root file, normally ‘C’ in windows, works better
Executable file: MapMaker.exp (command prompt)
Citation:
Lander E.S., P. Green, J. Abrahamson, A. Barlow, M.J. Daly, S.E. Lincoln and L.
Newburg (1987). MAPMAKER: An interactive computer package for constructing
primary genetic linkage maps of experimental and natural populations.
Genomics 1(2):174-181.
Sample Data File for Linkage Mapping
Step 1. Go to
https://plant-breeding-genomics.extension.org/mapmaker-tutorial/
Step 2. Download “mapmakersampledataset.xls” (from
attachments), copy and paste
“mapmakersampledataset.txt” to notepad (save as same)
Save in folder where you would want to store the output
files
Data source:
Scott Wolfe (2012). MapMaker Tutorial. Web accessed: Aug 26, 2020 from:
https://plant-breeding-genomics.extension.org/mapmaker-tutorial/
Sample Data File for Linkage Mapping
Mapping file information a:
Total number of markers: 27
Mapping population: F2 intercross (Parent A x Parent B)
Total number of individuals: 104
Marker symbol:
- 1 = Parent A (homozygous for parent A alleles)
- 2 = Heterozygous (both parent A and parent B alleles)
- 3 = Parent B (homozygous for parent B alleles)
- 4 = Not homozygous for parent A
- 5 = Not homozygous for parent B
a https://pbgworks.org/sites/pbgworks.org/files/mapmakersampletextfile.txt
Sample Data File for Linkage Mapping
Step 3. Check input file format:
Source: https://pbgworks.org/sites/pbgworks.org/files/mapmakersampletextfile.txt
Type of Population
Population
Size
Number of
Markers
Defaults
Genotype Score
ScoresMarker Names
Start MapMaker
Step 4. Double click MapMaker.exe and run the program
Set Working Directory
Step 5. Change directory (cd command) to folder where your input
file mapmakersampledataset.txt is located
Upload Input File
Step 6. Upload the input file using prepare command
Here, prepare mapmakersampledataset.txt
Saving work
Step 7. Save occasionally to avoid loss of work. Use photo
command. Here, saved as “output1.out”.
Specify data
Step 8. Specify data to be used using sequence command. Here all
marker data is selected
Grouping
Step 9. Build preliminary linkage groups using group command.
Default thresholds are LOD = 3 and max. rf = 50
Two groups with
14 and 13
markers; no
unlinked markers
Grouping
Step 10. Check at different LOD and max. rf values. Here,
two groups remain unchanged at higher values.
One unlinked at
LOD =7 and max.
rf. = 30 (very
stringent values).
Back to original
grouping
Working on Group 1
Step 11. Specify the group (use seq) to start working on that
group. Here, start with the first group identified as group 1.
Ordering Markers in Group 1
Step 12. Linear order of markers in a specified group can be
obtained using order command
Automatic
Ordering steps:
1. Finds most
informative subset
and map them
2. Adds remaining
markers individually
Ordering Markers in Group 1
Step 12. Linear order of markers in a specified group can be
obtained using order command
Automatic
Ordering steps:
1. Finds most
informative subset
and map them
2. Adds remaining
markers individually
3. Tries unmapped
ones at lower
threshold
Ordering Markers in Group 1
Step 12. Linear order of markers in a specified group can be
obtained using order command
Automatic
Ordering steps:
1. Finds most
informative subset
and maps them
2. Adds remaining
markers individually
3. Tries unmapped
ones at lower
threshold
4. Reports markers
that do not fit
uniquely
Add Remaining Markers to Group 1
Step 13. First, seq order1 (best fitted group 1 markers). Then, add
remaining markers with try command. Remember, different
original subset could lead to different unassigned markers
Adding unassigned markers:
1. Try remaining markers. Start with
first one (marker 10 in this case)
2. Marker 10 best fits 3rd position
Update Group 1
Step 14. Make new sequence with additional marker at best fit
position, add remaining markers, and build final sequence
Adding unassigned markers:
3. Make new sequence with marker
10 added to 3rd position
4. Try other unassigned markers
sequentially
5. Make updated sequence
Finalize Linkage Group 1
Step 15. Finally, map command is used to build genetic linkage
map of the first group.
Linkage Group 2
Step 16. Repeat Steps 11 to 15 to build remaining linkage groups
(here, second linkage group)
Genetic Linkage Maps
MapChart a used for graphical presentation of genetic linkage map
a Source: https://www.wageningenur.nl/en/show/Mapchart.htm

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Linkage mapping lab

  • 1. Lab Manual Sameer Khanal available at: https://www.slideshare.net/SameerKhanal1
  • 2. Linkage Mapping using MapMaker Software download site: http://research.franklin.uga.edu/devoslab/scripts-used-genetic-mapping Latest version: MapMaker/EXP 3.0b January 1993 Destination: closest to root file, normally ‘C’ in windows, works better Executable file: MapMaker.exp (command prompt) Citation: Lander E.S., P. Green, J. Abrahamson, A. Barlow, M.J. Daly, S.E. Lincoln and L. Newburg (1987). MAPMAKER: An interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1(2):174-181.
  • 3. Sample Data File for Linkage Mapping Step 1. Go to https://plant-breeding-genomics.extension.org/mapmaker-tutorial/ Step 2. Download “mapmakersampledataset.xls” (from attachments), copy and paste “mapmakersampledataset.txt” to notepad (save as same) Save in folder where you would want to store the output files Data source: Scott Wolfe (2012). MapMaker Tutorial. Web accessed: Aug 26, 2020 from: https://plant-breeding-genomics.extension.org/mapmaker-tutorial/
  • 4. Sample Data File for Linkage Mapping Mapping file information a: Total number of markers: 27 Mapping population: F2 intercross (Parent A x Parent B) Total number of individuals: 104 Marker symbol: - 1 = Parent A (homozygous for parent A alleles) - 2 = Heterozygous (both parent A and parent B alleles) - 3 = Parent B (homozygous for parent B alleles) - 4 = Not homozygous for parent A - 5 = Not homozygous for parent B a https://pbgworks.org/sites/pbgworks.org/files/mapmakersampletextfile.txt
  • 5. Sample Data File for Linkage Mapping Step 3. Check input file format: Source: https://pbgworks.org/sites/pbgworks.org/files/mapmakersampletextfile.txt Type of Population Population Size Number of Markers Defaults Genotype Score ScoresMarker Names
  • 6. Start MapMaker Step 4. Double click MapMaker.exe and run the program
  • 7. Set Working Directory Step 5. Change directory (cd command) to folder where your input file mapmakersampledataset.txt is located
  • 8. Upload Input File Step 6. Upload the input file using prepare command Here, prepare mapmakersampledataset.txt
  • 9. Saving work Step 7. Save occasionally to avoid loss of work. Use photo command. Here, saved as “output1.out”.
  • 10. Specify data Step 8. Specify data to be used using sequence command. Here all marker data is selected
  • 11. Grouping Step 9. Build preliminary linkage groups using group command. Default thresholds are LOD = 3 and max. rf = 50 Two groups with 14 and 13 markers; no unlinked markers
  • 12. Grouping Step 10. Check at different LOD and max. rf values. Here, two groups remain unchanged at higher values. One unlinked at LOD =7 and max. rf. = 30 (very stringent values). Back to original grouping
  • 13. Working on Group 1 Step 11. Specify the group (use seq) to start working on that group. Here, start with the first group identified as group 1.
  • 14. Ordering Markers in Group 1 Step 12. Linear order of markers in a specified group can be obtained using order command Automatic Ordering steps: 1. Finds most informative subset and map them 2. Adds remaining markers individually
  • 15. Ordering Markers in Group 1 Step 12. Linear order of markers in a specified group can be obtained using order command Automatic Ordering steps: 1. Finds most informative subset and map them 2. Adds remaining markers individually 3. Tries unmapped ones at lower threshold
  • 16. Ordering Markers in Group 1 Step 12. Linear order of markers in a specified group can be obtained using order command Automatic Ordering steps: 1. Finds most informative subset and maps them 2. Adds remaining markers individually 3. Tries unmapped ones at lower threshold 4. Reports markers that do not fit uniquely
  • 17. Add Remaining Markers to Group 1 Step 13. First, seq order1 (best fitted group 1 markers). Then, add remaining markers with try command. Remember, different original subset could lead to different unassigned markers Adding unassigned markers: 1. Try remaining markers. Start with first one (marker 10 in this case) 2. Marker 10 best fits 3rd position
  • 18. Update Group 1 Step 14. Make new sequence with additional marker at best fit position, add remaining markers, and build final sequence Adding unassigned markers: 3. Make new sequence with marker 10 added to 3rd position 4. Try other unassigned markers sequentially 5. Make updated sequence
  • 19. Finalize Linkage Group 1 Step 15. Finally, map command is used to build genetic linkage map of the first group.
  • 20. Linkage Group 2 Step 16. Repeat Steps 11 to 15 to build remaining linkage groups (here, second linkage group)
  • 21. Genetic Linkage Maps MapChart a used for graphical presentation of genetic linkage map a Source: https://www.wageningenur.nl/en/show/Mapchart.htm