PMID- 31583655
OWN - NLM
STAT- MEDLINE
DCOM- 20201211
LR  - 20201214
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2061
DP  - 2020
TI  - Preparing Maize Synaptonemal Complex Spreads and Sequential Immunofluorescence 
      and Fluorescence In Situ Hybridization.
PG  - 79-115
LID - 10.1007/978-1-4939-9818-0_8 [doi]
AB  - Immunofluorescence and fluorescence in situ hybridization (FISH) can be used to 
      locate specific proteins and DNA sequences, respectively, in chromosomes by light 
      microscopy. Here we describe sequential use of these techniques on spreads of 
      maize synaptonemal complexes (SCs) to determine whether crossing over can occur 
      in knob heterochromatin. We used antibodies to AFD1, an SC protein, and MLH1, a 
      class I (interference-sensitive) crossover protein found in most recombination 
      nodules (RNs) to identify crossovers (COs) along SCs. Next, we used FISH to 
      localize a 180 bp knob-specific tandem repeat. Combining immunofluorescence and 
      FISH images of the same SC spreads showed that heterochromatic knobs do not 
      prohibit class I COs. This technique is broadly applicable to investigations of 
      plant prophase I chromosomes where meiotic recombination takes place.
FAU - Stack, Stephen M
AU  - Stack SM
AD  - Department of Biology, Colorado State University, Fort Collins, CO, USA. 
      Stephen.Stack@colostate.edu.
FAU - Shearer, Lindsay A
AU  - Shearer LA
AD  - Department of Biology, Colorado State University, Fort Collins, CO, USA.
FAU - Lohmiller, Leslie D
AU  - Lohmiller LD
AD  - Department of Biology, Colorado State University, Fort Collins, CO, USA.
FAU - Anderson, Lorinda K
AU  - Anderson LK
AD  - Department of Biology, Colorado State University, Fort Collins, CO, USA.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Heterochromatin)
SB  - IM
MH  - Data Analysis
MH  - Fluorescent Antibody Technique
MH  - Heterochromatin
MH  - Image Processing, Computer-Assisted
MH  - *In Situ Hybridization, Fluorescence/methods
MH  - Recombination, Genetic
MH  - *Synaptonemal Complex
MH  - Zea mays/*genetics
OTO - NOTNLM
OT  - AFD1
OT  - Crossing over
OT  - Fluorescence in situ hybridization
OT  - Heterochromatin
OT  - Immunofluorescence
OT  - Knobs
OT  - MLH1
OT  - Maize
OT  - Recombination nodules
OT  - Synaptonemal complex
EDAT- 2019/10/05 06:00
MHDA- 2020/12/15 06:00
CRDT- 2019/10/05 06:00
PHST- 2019/10/05 06:00 [entrez]
PHST- 2019/10/05 06:00 [pubmed]
PHST- 2020/12/15 06:00 [medline]
AID - 10.1007/978-1-4939-9818-0_8 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2020;2061:79-115. doi: 10.1007/978-1-4939-9818-0_8.