mRNA levels of FKBP5 transcripts in Cohort 3 were measured using qRT-PCR. 40 mg of grey matter was dissected from each tissue block. RNA was extracted and RIN quantified as for RNA sequencing in Cohort 1. cDNA was synthesised from 500 ng of RNA using the Maxima H Minus Reverse Transcriptase system (ThermoFisher Scientific). PCR conditions were set to 94 °C for 5 min, 40 cycles of 94 °C for 30 s, 60 °C for 30 s, 72 °C for 30 s, and 72 °C for 7 min after the last cycle. Primer pairs were designed to amplify the unique junctions for total FKBP5 gex (Online Resource, Supplementary Table 7), using customised TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) and the Lightcycler 480 (Roche, Basel, Switzerland). Gex levels of FKBP5 was normalised to geometric means of the constitutively expressed genes β-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Samples were measured in quadruplicate and averaged.
Maxima h minus reverse transcriptase system
Manufactured by Thermo Fisher Scientific
The Maxima H Minus Reverse Transcriptase system is a reverse transcriptase enzyme designed for the synthesis of first-strand cDNA from RNA templates. It is suitable for use in a variety of RT-PCR applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using maxima h minus reverse transcriptase system
1
Quantification of FKBP5 mRNA Levels
2
Quantitative FKBP5 mRNA Expression Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
9. Quantitative Real-Time PCR mRNA levels of FKBP5 transcripts in Series 3 were measured using qRT-PCR. 40mg of grey matter was dissected from each tissue block. RNA was extracted and RIN quantified as for RNA sequencing in Series 1. cDNA was synthesised from 500ng of RNA using the Maxima H Minus Reverse
Transcriptase system (ThermoFisher Scientific). PCR conditions were set to 94°C for 5min, 40 cycles of 94°C for 30 s, 60°C for 30 s, 72°C for 30 s, and 72°C for 7min after the last cycle. Primer pairs were designed to amplify the unique junctions for total FKBP5 gex (Supplemental Table 6), using customized TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) and the Lightcycler 480 (Roche, Basel, Switzerland). Gex levels of FKBP5 was normalized to geometric means of the constitutively expressed genes β-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Samples were measured in quadruplicate and averaged. Normalised mRNA levels were then checked for normal distribution and correlated with age using spearman correlations in R.
Transcriptase system (ThermoFisher Scientific). PCR conditions were set to 94°C for 5min, 40 cycles of 94°C for 30 s, 60°C for 30 s, 72°C for 30 s, and 72°C for 7min after the last cycle. Primer pairs were designed to amplify the unique junctions for total FKBP5 gex (Supplemental Table 6), using customized TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) and the Lightcycler 480 (Roche, Basel, Switzerland). Gex levels of FKBP5 was normalized to geometric means of the constitutively expressed genes β-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Samples were measured in quadruplicate and averaged. Normalised mRNA levels were then checked for normal distribution and correlated with age using spearman correlations in R.
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!