A Fast Air-dry Dropping Chromosome Preparation Method Suitable for FISH in Plants

作者：Aliyeva Schnorr Lala; Ma Lu; Houben Andreas^*

来源：Jove-Journal of Visualized Experiments, 2015, (106): e53470.

DOI：10.3791/53470

• Plant Biology
• Issue 106
• chromosome preparation
• fluorescence in situ hybridization
• plants
• single-copy FISH
• mitotic metaphase
• relative humidity

Preparation of chromosome spreads is a prerequisite for the successful performance of fluorescence in situ hybridization (FISH). Preparation of high quality plant chromosome spreads is challenging due to the rigid cell wall. One of the approved methods for the preparation of plant chromosomes is a so-called drop preparation, also known as drop-spreading or air-drying technique. Here, we present a protocol for the fast preparation of mitotic chromosome spreads suitable for the FISH detection of single and high copy DNA probes. This method is an improved variant of the air-dry drop method performed under a relative humidity of 50%-55%. This protocol comprises a reduced number of washing steps making its application easy, efficient and reproducible. Obvious benefits of this approach are well-spread, undamaged and numerous metaphase chromosomes serving as a perfect prerequisite for successful FISH analysis. Using this protocol we obtained high-quality chromosome spreads and reproducible FISH results for Hordeum vulgare, H. bulbosum, H. marinum, H. murinum, H. pubiflorum and Secale cereale.

• 出版日期2015-12