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 PCR
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.

Polymerase Chain Reaction (PCR)

TAAATATTGCAATTTTCACGTGCAGGACGTGCGTGGACAACATCAAATCGATGGTTCCCTACTGTCCTCC ACGCTTCTTTGAGTTTGATCAAGAGCTCTACACAGGTATAATAGCATTTGCATGCACACATCATCACAAT GGGCACCGATTAATTCTCAACCTTCATCTTGCTTCAATTTTGTGCAACAGCAGAGTATGATGCTTCGAGG TTGATGAGGAAGCAGTTGGCAGAGAAGCGCGAGATGCTCTTGGAGATGGAGAGGCGGCGCGCCACAGTTC GGAGGCTTGAATCCATGCCGCCACAGTTTGCATACTTCGATTGCAGCATTGAGGACGCCAGTCCCCTCCA TTCTCTCCAGGATGGTCTGTGTGTTGTGTCATTCATTGGTCATTGCCAGAATTATACCTCTAGTTGTTCC ATTTCTCCCTGGTTGAAGTTTGTTTAGTGGCTCAAGGATTATCTACTTGATTATAGTGATAAATTAAATA ATTTAAGTAATGAACTTAATGATTAGGTTAATAAGTGCAGTAGAAGAACGGTGTTTCTTACCAAAACATA TTTTTTGAAGCGTGGAGAAATGTTTTAATAGTATAGTGAATACTGAATAGTGCACGGCCAAATATCCTGA TGGTTTTATGCGGCGTTTTTTTCCTGTGAACAGATTCGAAACAACTCGACCTTATGAATCCATCCCTTGC ATCCCCTGATCCACTGGAGATCGTTTCAAACAGCCAAGCTCCTCCTACACAGGCAGGCAACATTTATGAT GACCATGAGAGGTACAAACATATTTGCAGCCAATTTCTTGCATATATGTTGCTACCTGACATTGAGCTAG TTCCTCTTTGTTCACTGGATTGCTTCATTTGATTTCTAACTGCAGTAACCAGATTGAACTGCTGCCTGAG AGTCCATTCGCCGCATCAGCGCCTGCTGGAAACAGCATATCCACAATTATATAGGAGGAAAAAAATTACA AAGAGACTTTTGGGCCTTGGGTTCGGAATTCAGGTGCCCCTTTAGGCATTTGATGGTAATTTGGTCCAAG AGTAACATAAGTATACACACACAGGAGGAGACAACAAAAAGCAACAATATTTTTGTTGTAGATAGAGAAG GAAGGCAGAAGAAAATCTCACTGGTCATAATTAGGAGGCACATCTTTTTTGAAAAAAAATAATTAGGAGG CACATAAACACATGGTGTTCATTTTATACTTTAGAAGAGGACAGAAACAACAGCTTACAAAAGAATCTCA CATGGTGCAGTAGCTAGCTTGCAAGAGGTAAGGACAGTTACAAAAGAATTACAATGGTGGAAACTTGAGA TATAGATGTGCAGTTTGACTATAATTAAGGAATAGGGGACAATGTGATATGCTAATTATACTGTCCTAGA ATAAGGCATGTTATTATTTACATATTTAGTGTTATCTTCTTTTTTTTAGGGTATAATGGCCTACTGGCCT TTACCTTTACAAGGGTGAAAAAAAAAAGCAGAACCTTTGCTGATATTCCTATTGACAAGATGAGCAGAGC GTGCCCATTTGTACTTATCATCAGTAATGGAGAAAATTGCCTTCGATGTTTATTTCCTGCTTTCTGTAAA TGATTTGTCTTGTTTTGATTGCATTGATATTTTGCAAGAGTAACAGATAAAAACTTTGAGTGCATCATTA GATCGCATTTTCACC

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 (max = 488 nm) and emits green light (max = 522 nm).

Stratagene MX3005p

Consolidated Report

Amplification Plots

Dissociation Curve

Assay
G1 G1 G1 G1 G1 G1 G3 G3 G3 G3 G3 G3 normalizer normalizer normalizer normalizer normalizer normalizer

Well Type
Calibrator Calibrator Calibrator Unknown (Stress) Unknown (Stress) Unknown (Stress) Calibrator Calibrator Calibrator Unknown (Stress) Unknown (Stress) Unknown (Stress) Calibrator Calibrator Calibrator Unknown (Stress) Unknown (Stress) Unknown (Stress)

Ct (dRn)
33.7 33.64 32.48 26.09 26.14 26.28 26.63 26.69 26.58 22.48 22.48 22.48 19 18.97 18.95 19.69 19.65 19.74

Assay
G1 G1 G3 G3 normalizer normalizer

Well Type
Calibrator Unknown (Stress) Calibrator Unknown (Stress) Calibrator Unknown (Stress)

Ct (dRn)
33.15 26.17 26.64 22.48 18.97 19.69

Relative Quantity Chart

qPCR Analysis