SYBR™ Green PCR Master Mix

PCR Quantitative real-time PCR - Mammalian DNA

Experiment
PCR Quantitative real-time PCR - Mammalian DNA
Product
SYBR™ Green PCR Master Mix from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.
Protocol tips
PCR plates, tubes and pipette tips should be UV sterilized for 20-30 mins

Publication protocol

RT-PCR
RNA was extracted using Trizol (Invitrogen) according to the manufacturer’s instructions. RNA (5μg) was used for cDNA synthesis utilizing the Superscript III strand synthesis system (Invitrogen). For qualitative analysis of POLA1 transcripts, the following primer sequences were utilized: primer A (exon 10, 5’-AAAGGGGCAGATGAGGAACAA-3’); primer X (exon 13a, 5’-TCTGACAGTGGTGATGAAAAG-3’); primer B (exon 15, 5’-ACAAG CGGTGGTGGACTGAC-3’). Quantitative real-time RT-PCR was performed using SYBR Green based detection (Invitrogen) and a Mastercycler (Eppendorf, Germany) as previously reported46. Experiments were performed using technical duplicates or triplicates, data were normalized to housekeeping genes and the relative abundance of transcripts was calculated by the comparative ΔΔCt method. All primers used for qRT-PCR are indicated in Supplementary Table 6.



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Discussion

Discussion

5 years ago

Author: Germany

What is the optimal concentration for primers in qPCR?

What is the optimal concentration for primers in qPCR? My total volume is 20μl per reaction.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing PCR Quantitative real-time PCR - Mammalian DNA using SYBR™ Green PCR Master Mix from Thermo Fisher Scientific.

Paper title
DNA polymerase-α regulates type I interferon activation through cytosolic RNA:DNA synthesis
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for SYBR™ Green PCR Master Mix below.

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