Young root tips collected from parental cuttings and from the putative amphihaploid plantlets were held in ice water for 20–24 h, then fixed in 3:1 ethanol:glacial acetic acid (v/v) at 4 °C for 24 h, and squashed in a drop of 45% acetic acid. Mitotic chromosome spreads were observed by phase contrast microscopy. The mitotic chromosomes of the putative amphihaploid were also subjected to genomic in situ hybridization (GISH) by probing with genomic DNA of C. zawadskii extracted from young leaves using the CTAB (cetyltrimethylammonium bromide) method (Stewart and Via 1993 (link)) and labeled with biotinylated 16-dUTP (Roche Ltd.). The GISH procedure followed Deng et al. (2010) (link).
Biotinylated 16 dutp
Manufactured by Roche
Biotinylated 16-dUTP is a nucleotide analog used in various molecular biology applications. It contains a biotin molecule attached to the 16th position of the deoxyuridine triphosphate (dUTP) structure. The biotin label allows for the detection and purification of nucleic acids containing the modified nucleotide.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using biotinylated 16 dutp
1
Chromosome Analysis of Amphihaploid Plantlets
2
Spatial Distribution of Apoptotic Cells After Ischemia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To clarify the spatial distribution of apoptotic cells after cerebral ischemia, we performed terminal deoxynucleotidyl transferase-mediated uridine 5′-triphosphate-biotin nick end labeling (TUNEL) staining as previously described (n = 8 each) [1 (link)]. Fixed sections were incubated with NeuroPore (Trevigen) for 30 min. They were placed in 1 × terminal deoxynucleotidyl transferase (TdT) buffer (Invitrogen) with a TdT enzyme (Invitrogen) and biotinylated 16-dUTP (Roche Diagnostics) at 37 °C for 90 min. The avidin–biotin technique was applied and then the nuclei were counterstained with methyl green solution for 2 min. For more precise confirmation about SR-FLIVO-FMNP probe tracking within the apoptotic cells, high resolution transmission electron microscopy (HRTEM) was recorded for the cells located at ischemic and non-ischemic lesions.
3
In Situ Apoptosis Detection by TUNEL Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptotic cells were detected by TUNNEL assay. 8 μm thickness coronal sections were put in 1x terminal deoxynucleotidyl transferase buffer (Invitrogen, Carlsbad, CA) for 30 min, then, they reacted with terminal deoxynucleotidyl transferase enzyme (Invitrogen) and biotinylated 16-dUTP (Roche Diagnostics, Indianapolis, IN) at 37 °C for 60 min. The slices were washed with 2x SSC (150 mol/ liter sodium chloride and 15 mol/liter sodium citrate, pH 7.4) for 15 min, and then washed with PBS for 15 mins two times again. Counterstaining nuclei with methyl green solution using avidin-biotin technique (46) .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!