RNA was purified from cell lines using the QIAgen miRNeasy kit (Valencia, CA) and RT-PCR was performed to amplify FGFR3-L and FGFR3-S using random hexamers (Thermo Fisher Scientific). 5’ and 3’ rapid amplification of cDNA ends (RACE) was performed as previously described8 (link). RACE primers were designed based on the reference sequence of FGFR3 provided by the National Center for Biotechnology Information (NCBI) and are listed in Supplemental Table S1 . RT-PCR products were ligated into pCR2.1-TOPO TA vector (Thermo Fisher Scientific) according to manufacturer’s instructions. Three to 4 clones per cell line were sequence verified (Sequetech, Mountain View, CA) and the consensus sequence of the full-length FGFR3-S variant was deposited in GenBank (Accession #MK542707 ).
Protocol detail
Identification and Characterization of FGFR3 Isoforms
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : George Washington University
Other organizations : University of Maryland Eastern Shore, University of Hawaii System, University of Michigan–Ann Arbor, National Institute of Dental and Craniofacial Research, Uniformed Services University of the Health Sciences
Top 5 similar protocols
Variable analysis
independent variables
- RNA purification using the QIAgen miRNeasy kit
- RT-PCR to amplify FGFR3-L and FGFR3-S using random hexamers
- 5' and 3' rapid amplification of cDNA ends (RACE)
- Expression levels of FGFR3-L and FGFR3-S
- Cell lines used for RNA purification
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!