The buccal cells donated by the subjects were collected in Resuspension Solution (GenElute Mammalian Genomic DNA Miniprep Kit, Sigma, Germany) with the use of sterile foam-tipped applicators (Puritan, USA). DNA was extracted from the buccal cells using a GenElute Mammalian Genomic DNA Miniprep Kit (Sigma, Germany) according to the manufacturer's protocol. All samples were genotyped in duplicate using an allelic discrimination assay on a StepOne Real-Time Polymerase Chain Reaction (RT-PCR) instrument (Applied Biosystems, USA) as previously described [19 (link)].
Stepone real time polymerase chain reaction rt pcr instrument
Manufactured by Thermo Fisher Scientific
Sourced in United States
The StepOne Real-Time Polymerase Chain Reaction (RT-PCR) instrument is a compact and versatile system designed for real-time PCR analysis. It is capable of performing quantitative gene expression analysis, genotyping, and copy number variation studies. The StepOne system utilizes a 96-well block format and is compatible with a wide range of commercially available PCR assays and reagents.
Automatically generated - may contain errors
5 protocols using stepone real time polymerase chain reaction rt pcr instrument
1
Buccal Cell DNA Extraction and Genotyping
2
Buccal Cell DNA Extraction and Genotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The buccal cells donated by the subjects were collected in Resuspension Solution (GenElute Mammalian Genomic DNA Miniprep Kit, Sigma, Germany) with the use of sterile foam-tipped applicators (Puritan, USA). DNA was extracted from the buccal cells using a GenElute Mammalian Genomic DNA Miniprep Kit (Sigma, Germany) according to the manufacturer’s protocol. All samples were genotyped in duplicate using an allelic discrimination assay on a StepOne Real-Time Polymerase Chain Reaction (RT-PCR) instrument (Applied Biosystems, USA) as previously described (Zarebska et al., 2014 (link)).
3
Genotyping of GSTP1 Polymorphism
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNA was extracted from buccal cells using a GenElute Mammalian Genomic DNA Miniprep Kit (Sigma, Germany) according to the manufacturer's protocol. All samples were genotyped using an allelic discrimination assay on a StepOne Real-Time Polymerase Chain Reaction (RT-PCR) instrument (Applied Biosystems, USA) with TaqMan probes. To discriminate GSTP1 A and G alleles (rs1695), a TaqMan Pre-Designed SNP Genotyping Assay was used (Applied Biosystems, USA) (assay ID: C___3237198_20), including primers and fluorescently labelled (FAM and VIC) MGB probes to detect both alleles. All sample were analysed in duplicate, and there was 100% agreement in genotype detection between samples.
4
Buccal Cell DNA Extraction and Genotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The buccal cells donated by the subjects were collected in Resuspension Solution (GenElute Mammalian Genomic DNA Miniprep Kit, Sigma, Germany) using sterile foam-tipped applicators (Puritan, USA). DNA was extracted from the buccal cells using a GenElute Mammalian Genomic DNA Miniprep Kit (Sigma, Germany) according to the manufacturer’s protocol. All samples were genotyped in duplicate using an allelic discrimination assay on a StepOne Real-Time Polymerase Chain Reaction (RT-PCR) instrument (Applied Biosystems, USA).
5
Genotyping of COL22A1 Variants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The buccal cells were collected using Copan FLOQSwabs (Copan Diagnostics, Inc., Murrieta, CA, USA) during post-surgery control. DNA was extracted using a GenElute Mammalian Genomic DNA Miniprep Kit (Sigma, Taufkirchen, Germany) in agreement with the manufacturer’s instructions. On a StepOne Real-Time Polymerase Chain Reaction (RT-PCR) instrument (Applied Biosystems, Bedford, MA, USA), all samples were genotyped in duplicate using the TaqMan® pre-designed SNP genotyping assays (C_176045196_10 for the COL22A1 rs11784270 and C_29400116_10 for the COL22A1 rs6577958, Applied Biosystems, Bedford, MA, USA) according to the protocol [12 (link)].
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!