Genomic DNA was extracted from the leaves of each petunia cultivar using the DNeasy plant Maxi kit (QIAGEN, Venlo, Netherlands), following the manufacturer’s instructions. Eco RI and Xba I were used to digest 10 μg of P. hybrida genomic DNA, which was then separated on a 0.9% agarose gel and transferred to a positively charged nylon membrane (Roche Applied Science, Basel, Switzerland). We used the coding region of eYGFPuv as the DNA probe in this analysis, and it was labelled using a PCR DIG probe synthesis kit (Roche Applied Science) and the primers 5′-ATGACAACCTTCAAAATCGAGTCC and 5′-CTACATGTCTCTTGGGGCGCTGTT. Hybridization signals were detected by chemiluminescence using CDP-Star (Roche Applied Science) as the substrate, and visualized with an imaging system (Molecular Imager VersaDoc™ MP 5000, Bio-Rad, Hercules, CA, USA).
Molecular imager versadoc mp 5000
Manufactured by Bio-Rad
Sourced in United States
The Molecular Imager VersaDoc™ MP 5000 is a laboratory imaging system designed for the capture and analysis of various types of gel and blot-based assays. It features a high-resolution camera and advanced optics to enable sensitive and quantitative detection of fluorescent, colorimetric, and chemiluminescent signals.
Automatically generated - may contain errors
Lab products found in correlation
Positively charged nylon membrane, by Roche (1 mentions)
Dneasy plant maxi kit, by Qiagen (1 mentions)
Pcr dig probe synthesis kit, by Roche (1 mentions)
Cdp star, by Roche (1 mentions)
Western breeze chemiluminescent kit, by Thermo Fisher Scientific (1 mentions)
Amicon ultra device, by Merck Group (1 mentions)
Cytobuster protein extraction reagent, by Merck Group (1 mentions)
2 protocols using molecular imager versadoc mp 5000
1
Petunia Genomic DNA Extraction and Analysis
2
Quantification of Phosphorylated Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were extracted using the CytoBuster™ Protein Extraction Reagent (Merck Millipore, Billerica, MA, USA) and concentrated or diafiltered with an Amicon Ultra device (Merck Millipore). Cell lysates were subjected to SDS-PAGE (12.5%), followed by western blotting. The target proteins were visualized using a WesternBreeze® Chemiluminescent Kit (Thermo Fisher Scientific, Waltham, MA, USA) and were detected using a Molecular Imager VersaDoc™ MP 5000 (Bio-Rad, Hercules, CA, USA). The band intensity of western blots was determined by densitometry with Image J and the mean value was obtained. The degrees of phosphorylation of signal proteins were expressed as the relative ratio of the amount of phosphorylated signal protein compared with the amount of the intact signal protein, i.e. phosphorylated NGF receptor/NGF receptor (P-NGFR/NGFR), phosphorylated Erk/Erk (P-Erk/Erk), phosphorylated Akt/Akt (P-Akt/Akt) or phosphorylated CaMKII/CaMKII (P-CaMKII/CaMKII).
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!