We extracted total RNAs from cultured cells or tumorous tissues using Tri-RNA extraction reagent (Sigma, St. Luis, MO, USA), and performed real-time quantitative PCR analyses according to a previously published procedure (57 (link)). For each sample, we used triplicates and used the comparative CT (ΔΔCT) as described by the manufacturer (Applied Biosystems/ Fisher Scientific). The relative amount of target (2ȒΔΔCT) was obtained by normalizing to an endogenous glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and relative to a calibrator. All primers and probes were purchased from Applied Biosystems/Fisher Scientific). RT-PCR was used to detect possible existence of Gli1 isoforms in the pancreatic cancer cell lines. Primers 5′-TGTTCAACTCGATGACCC-3′ & 5′-GTCATGGGGACCACAAGG-3′ were used to detect wild type Gli1 (500bp) and truncated Gli1 (tGli1) (377bp). Primers 5′-GGCATCCGACAGAGGTGAGATGGAC-3’ and 5′-GAGCCCAGCGCCCAGACAGA-3’ or 5′-CTGTCTCAGGGAACCGTGGGTCTTTGT-3’ were used to detect full-length Gli1 or Gli1ΔN in pancreatic cancer cell lines.
Tri rna extraction reagent
Manufactured by Merck Group
Sourced in United States
Tri-RNA extraction reagent is a laboratory product manufactured by Merck Group. It is designed for the isolation and purification of total RNA from a variety of biological samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using tri rna extraction reagent
1
Quantitative PCR and RT-PCR Analysis of Gli1 Isoforms in Pancreatic Cancer
2
Quantitative PCR Analysis of GLI1 Isoforms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We extracted total RNAs from cultured cells or tumorous tissues using Tri-RNA extraction reagent (Sigma, St. Luis, MO, USA), and performed real-time quantitative PCR analyses according to a previously published procedure [57 (link)]. For each sample, we used triplicates and used the comparative CT (ΔΔCT) as described by the manufacturer (Applied Biosystems/ Fisher Scientific). The relative amount of target (2−ΔΔCT) was obtained by normalizing to an endogenous glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and relative to a calibrator. All primers and probes were purchased from Applied Biosystems/Fisher Scientific). RT-PCR was used to detect possible existence of GLI1 isoforms in the pancreatic cancer cell lines. Primers 5′-TGTTCAACTCGATGACCC-3′ and 5′-GTCATGGGGACCACAAGG-3′ were used to detect wild type GLI1 (500 bp) and truncated GLI1 (tGLI1) (377 bp). Primers 5′-GGCATCCGACAGAGGTGAGATGGAC-3′ and 5′-GAGCCCAGCGCCCAGACAGA-3′ or 5′-CTGTCTCAGGGAACCGTGGGTCTTTGT-3′ were used to detect full-length GLI1 or GLI1ΔN in pancreatic cancer cell lines.
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!