Real Time PCR

Transcription

The process of creating an equivalent RNA copy of a sequence of DNA.

mRNA processing

Reverse transcriptase, also known as RNA-dependent DNA polymerase, is a DNA polymerase enzyme that transcribes single-stranded RNA into double-stranded DNA.

Tools for Gene Expression

Northern analysis

Quantitative PCR

Dot/Slot blot analysis

Microarray

Real-time polymerase chain reaction, also called quantitative real time polymerase chain reaction (Q-PCR/qPCR/qrt-PCR) or kinetic polymerase chain reaction (KPCR), is a laboratory technique based on the PCR, which is used to amplify and simultaneously quantify a targeted DNA molecule.

Real-time PCR

Polymerase Chain Reaction (PCR)

TAAATATTGCAATTTTCACGTGCAGGACGTGCGTGGACAACATCAAATCGATGGTTCCCTACTGTCCTCCACGCTTCTTTGAGTTTGATCAAGAGCTCTACACAGGTATAATAGCATTTGCATGCACACATCATCACAATGGGCACCGATTAATTCTCAACCTTCATCTTGCTTCAATTTTGTGCAACAGCAGAGTATGATGCTTCGAGGTTGATGAGGAAGCAGTTGGCAGAGAAGCGCGAGATGCTCTTGGAGATGGAGAGGCGGCGCGCCACAGTTCGGAGGCTTGAATCCATGCCGCCACAGTTTGCATACTTCGATTGCAGCATTGAGGACGCCAGTCCCCTCCATTCTCTCCAGGATGGTCTGTGTGTTGTGTCATTCATTGGTCATTGCCAGAATTATACCTCTAGTTGTTCCATTTCTCCCTGGTTGAAGTTTGTTTAGTGGCTCAAGGATTATCTACTTGATTATAGTGATAAATTAAATAATTTAAGTAATGAACTTAATGATTAGGTTAATAAGTGCAGTAGAAGAACGGTGTTTCTTACCAAAACATATTTTTTGAAGCGTGGAGAAATGTTTTAATAGTATAGTGAATACTGAATAGTGCACGGCCAAATATCCTGATGGTTTTATGCGGCGTTTTTTTCCTGTGAACAGATTCGAAACAACTCGACCTTATGAATCCATCCCTTGCATCCCCTGATCCACTGGAGATCGTTTCAAACAGCCAAGCTCCTCCTACACAGGCAGGCAACATTTATGATGACCATGAGAGGTACAAACATATTTGCAGCCAATTTCTTGCATATATGTTGCTACCTGACATTGAGCTAGTTCCTCTTTGTTCACTGGATTGCTTCATTTGATTTCTAACTGCAGTAACCAGATTGAACTGCTGCCTGAGAGTCCATTCGCCGCATCAGCGCCTGCTGGAAACAGCATATCCACAATTATATAGGAGGAAAAAAATTACAAAGAGACTTTTGGGCCTTGGGTTCGGAATTCAGGTGCCCCTTTAGGCATTTGATGGTAATTTGGTCCAAGAGTAACATAAGTATACACACACAGGAGGAGACAACAAAAAGCAACAATATTTTTGTTGTAGATAGAGAAGGAAGGCAGAAGAAAATCTCACTGGTCATAATTAGGAGGCACATCTTTTTTGAAAAAAAATAATTAGGAGGCACATAAACACATGGTGTTCATTTTATACTTTAGAAGAGGACAGAAACAACAGCTTACAAAAGAATCTCACATGGTGCAGTAGCTAGCTTGCAAGAGGTAAGGACAGTTACAAAAGAATTACAATGGTGGAAACTTGAGATATAGATGTGCAGTTTGACTATAATTAAGGAATAGGGGACAATGTGATATGCTAATTATACTGTCCTAGAATAAGGCATGTTATTATTTACATATTTAGTGTTATCTTCTTTTTTTTAGGGTATAATGGCCTACTGGCCTTTACCTTTACAAGGGTGAAAAAAAAAAGCAGAACCTTTGCTGATATTCCTATTGACAAGATGAGCAGAGCGTGCCCATTTGTACTTATCATCAGTAATGGAGAAAATTGCCTTCGATGTTTATTTCCTGCTTTCTGTAAATGATTTGTCTTGTTTTGATTGCATTGATATTTTGCAAGAGTAACAGATAAAAACTTTGAGTGCATCATTAGATCGCATTTTCACC

Molecular Beacons for qPCR

TaqMan probe

Scorpions Probes

SYBR Green probe for qPCR

SYBR Green I (SG) is an asymmetrical cyanine dye used as a nucleic acid stain.DNA-dye-complex absorbs blue light (λmax = 488 nm) and emits green light (λmax = 522 nm).

Stratagene MX3005p

Consolidated Report

Amplification Plots

Dissociation Curve

Assay Well Type Ct (dRn)G1 Calibrator 33.7

G1 Calibrator 33.64

G1 Calibrator 32.48

G1 Unknown (Stress) 26.09

G1 Unknown (Stress) 26.14

G1 Unknown (Stress) 26.28

G3 Calibrator 26.63

G3 Calibrator 26.69

G3 Calibrator 26.58

G3 Unknown (Stress) 22.48

G3 Unknown (Stress) 22.48

G3 Unknown (Stress) 22.48

normalizer Calibrator 19

normalizer Calibrator 18.97

normalizer Calibrator 18.95

normalizer Unknown (Stress) 19.69

normalizer Unknown (Stress) 19.65

normalizer Unknown (Stress) 19.74

Assay Well Type Ct (dRn)

G1 Calibrator 33.15

G1 Unknown (Stress) 26.17

G3 Calibrator 26.64

G3 Unknown (Stress) 22.48

normalizer Calibrator 18.97

normalizer Unknown (Stress) 19.69

Relative Quantity Chart

qPCR Analysis