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Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

Posted on June 29, 2021April 29, 2021 by Leah

Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control.
The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data.

 

Pig Adrenal, Whole Total RNA*

PR-501 Zyagen 0.05mg
EUR 235

Pig Adrenal, Whole Total Protein

PT-501 Zyagen 0.5mg
EUR 153

Rat Adrenal Whole tissue lysate

RAL-1470 Alpha Diagnostics 100ug
EUR 255.6

Mouse Adrenal Whole tissue lysate

MAL-1417 Alpha Diagnostics 100ug
EUR 255.6

Bovine Eye, Whole cDNA

BD-106 Zyagen 30 reactions
EUR 243

Human Normal Adrenal Gland Whole tissue lysate

HALX-11050 Alpha Diagnostics 1 mg
EUR 927.6

Monkey (Cyno.) Normal Adrenal Gland Whole tissue lysate

MCL-1160 Alpha Diagnostics 1 mg
EUR 927.6

PC-12 (Rat adrenal pheochromocytoma) Whole Cell Lysate

E45O00018 EnoGene each
EUR 395

Monkey (Rhesus) Normal Adrenal Gland Whole tissue lysate

MRL-1267 Alpha Diagnostics 1 mg
EUR 927.6

Adrenal Gland, Bovine

MBS639367-1Each MyBiosource 1Each
EUR 510

Adrenal Gland, Bovine

MBS639367-5x1Each MyBiosource 5x1Each
EUR 2080

Dog Adrenal cDNA*

DD-501 Zyagen 30 Reactions
EUR 319

Cat Adrenal cDNA

FD-501 Zyagen 30 Reactions
EUR 319

Rat Adrenal cDNA *

RD-501 Zyagen 30 reactions
EUR 280

Sheep Adrenal cDNA*

SD-501 Zyagen 30 reactions
EUR 280

57101-2 BOVINE ADRENAL 50 EA*

57101-2 Pel-Freez 50 Each
EUR 917

Rabbit Adrenal cDNA

TD-501 Zyagen 30 reactions
EUR 243

Bovine Adrenal, Cortex Total RNA*

BR-506 Zyagen 0.05mg
EUR 235

Hamster Adrenal cDNA

AD-501 Zyagen 30 reactions
EUR 243

MiniPig Adrenal cDNA

ND-501 Zyagen 30 reactions
EUR 407

Bovine Adrenal, Medulla Total RNA*

BR-507 Zyagen 0.05mg
EUR 235

Bovine Adrenal, Cortex Total Protein

BT-506 Zyagen 0.5mg
EUR 153

Bovine Adrenal, Medulla Total Protein

BT-507 Zyagen 0.5mg
EUR 153

Bovine Adrenal Medulla 22P (BAM-22P)

MBS633930-1mg MyBiosource 1mg
EUR 610

Bovine Adrenal Medulla 22P (BAM-22P)

MBS633930-5x1mg MyBiosource 5x1mg
EUR 2600

Bovine Adrenal Medulla 18P (BAM-18P)

MBS658691-1mg MyBiosource 1mg
EUR 610

Bovine Adrenal Medulla 18P (BAM-18P)

MBS658691-5x1mg MyBiosource 5x1mg
EUR 2600

Bovine Adrenal Medulla 12P (BAM-12P)

MBS659233-1mg MyBiosource 1mg
EUR 610

Bovine Adrenal Medulla 12P (BAM-12P)

MBS659233-5x1mg MyBiosource 5x1mg
EUR 2600

Mouse CD1 Adrenal cDNA *

MD-501 Zyagen 30 reactions
EUR 280

Mouse BLC Adrenal cDNA *

MD-501-BLC Zyagen 30 reactions
EUR 319

Mouse C57 Adrenal cDNA *

MD-501-C57 Zyagen 30 reactions
EUR 319

Guinea Pig Adrenal cDNA

GD-501 Zyagen 30 reactions
EUR 243

Bovine Adrenal Microvascular Endothelial Cells

B1013 Alphabioregen 1ml frozen Vial
EUR 0.2

Bovine Adrenal Microvascular Endothelial Cells

cAP-b0008 Angio Proteomie 1Frozen Vial
EUR 742.5

Monkey Adrenal cDNA, Rhesus

UD-501 Zyagen 30 reactions
EUR 446

Monkey Adrenal cDNA, Cynomolgus

KD-501 Zyagen 30 reactions
EUR 446

BAM-12P, Bovine Adrenal Medulla Docosapeptide

5-00748 CHI Scientific 4 x 1mg
EUR 120.12
Description: Tyr-Gly-Gly-Phe-Met-Arg-Arg-Val-Gly-Arg-Pro-Glu

Bovine adrenal cortex antibody,ACA ELISA kit

E01A79939 BlueGene 96T
EUR 700
Description: ELISA

Bovine adrenal cortex antibody,ACA ELISA kit

E11A2028-48wellsplate BlueGene 48 wells plate
EUR 280

Bovine adrenal cortex antibody,ACA ELISA kit

E11A2028-96wellsplate BlueGene 96 wells plate
EUR 405

Bovine adrenal cortex antibody, ACA ELISA Kit

MBS7239599-10x96StripWells MyBiosource 10x96-Strip-Wells
EUR 5685

Bovine adrenal cortex antibody, ACA ELISA Kit

MBS7239599-48StripWells MyBiosource 48-Strip-Wells
EUR 485

Bovine adrenal cortex antibody, ACA ELISA Kit

MBS7239599-5x96StripWells MyBiosource 5x96-Strip-Wells
EUR 3020

Bovine adrenal cortex antibody, ACA ELISA Kit

MBS7239599-96StripWells MyBiosource 96-Strip-Wells
EUR 690

Bovine Adrenal Medulla 12P (BAM-12P) [7-12]

MBS658494-1mg MyBiosource 1mg
EUR 610

Bovine Adrenal Medulla 12P (BAM-12P) [7-12]

MBS658494-5x1mg MyBiosource 5x1mg
EUR 2600

cDNA - Human Tumor Tissue: Adrenal

C1235004-10 Biochain 10 reactions
EUR 221.2

Human Adrenal cDNA-Oligo-dT

HD-501 Zyagen 30 reactions
EUR 280

Rat Adrenal cDNA-Random Primer *

RD-501-RH Zyagen 30 reactions
EUR 280

Human Adrenal cDNA-Random Primer

HD-501-HR Zyagen 30 reactions
EUR 280

Innovative Grade US Origin Bovine Gland Adrenal

IGBOGLARFX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGBOGLARFX2 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGBOGLARPX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGBOGLARPX2 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGBOGLARSX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGBOGLARSX2 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGBOGLARZX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGBOGLARZX2 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGCLFGLARFX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGCLFGLARFX2 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGCLFGLARPX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGCLFGLARPX2 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGCLFGLARSX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGCLFGLARSX2 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGCLFGLARZX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Innovative Grade US Origin Bovine Gland Adrenal

IGCLFGLARZX2 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal

Rat WS Adrenal cDNA-Oligo-dT *

RD-501-WS Zyagen 30 reactions
EUR 280

cDNA - Human Adult Normal Tissue: Adrenal

C1234004 Biochain 40 reactions
EUR 311.5

Mouse C57 Adrenal cDNA-Random Primer *

MD-501-C57-RH Zyagen 30 reactions
EUR 319

Mouse CD1 Adrenal cDNA-Random Primer *

MD-501-HR Zyagen 30 reactions
EUR 280

Innovative Grade US Origin Bovine Gland Adrenal Cortex

IGBOGLARCFX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal Cortex

Innovative Grade US Origin Bovine Gland Adrenal Cortex

IGBOGLARCPX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal Cortex

Innovative Grade US Origin Bovine Gland Adrenal Cortex

IGBOGLARCSX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal Cortex

Innovative Grade US Origin Bovine Gland Adrenal Cortex

IGBOGLARCZX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal Cortex

Innovative Grade US Origin Bovine Gland Adrenal Cortex

IGCLFGLARCFX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal Cortex

Innovative Grade US Origin Bovine Gland Adrenal Cortex

IGCLFGLARCPX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal Cortex

Innovative Grade US Origin Bovine Gland Adrenal Cortex

IGCLFGLARCSX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal Cortex

Innovative Grade US Origin Bovine Gland Adrenal Cortex

IGCLFGLARCZX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal Cortex

cDNA - Monkey (Rhesus) Normal Tissue: Adrenal

C1534004 Biochain 40 reactions
EUR 531.3

Innovative Grade US Origin Bovine Gland Adrenal Medulla

IGBOGLARMFX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal Medulla

Innovative Grade US Origin Bovine Gland Adrenal Medulla

IGBOGLARMPX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal Medulla

Innovative Grade US Origin Bovine Gland Adrenal Medulla

IGBOGLARMSX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal Medulla

Innovative Grade US Origin Bovine Gland Adrenal Medulla

IGBOGLARMZX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal Medulla

Innovative Grade US Origin Bovine Gland Adrenal Medulla

IGCLFGLARMFX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal Medulla

Innovative Grade US Origin Bovine Gland Adrenal Medulla

IGCLFGLARMPX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal Medulla

Innovative Grade US Origin Bovine Gland Adrenal Medulla

IGCLFGLARMSX1 Innovative research each
EUR 210
Description: Innovative Grade US Origin Bovine Gland Adrenal Medulla

Innovative Grade US Origin Bovine Gland Adrenal Medulla

IGCLFGLARMZX1 Innovative research each
EUR 133
Description: Innovative Grade US Origin Bovine Gland Adrenal Medulla

Monkey Adrenal cDNA-Random Primer, Rhesus

UD-501-RH Zyagen 30 reactions
EUR 446

cDNA - Monkey (Cynomolgus) Normal Tissue: Adrenal

C1534004-Cy Biochain 40 reactions
EUR 531.3

Monkey Adrenal cDNA-Random Primer, Cynomolgus

KD-501-RH Zyagen 30 reactions
EUR 446

Peptide E / 3200-Dalton Adrenal Enkephalin-Containing Peptide (Bovine)

024-57 PHOENIX PEPTIDE 200 μg
EUR 86.4

Tissue cDNA, First Strand, Human Tumor, Adrenal, BioGenomics

MBS652063-10Tests MyBiosource 10Tests
EUR 570

Tissue cDNA, First Strand, Human Tumor, Adrenal, BioGenomics

MBS652063-5x10Tests MyBiosource 5x10Tests
EUR 2345

BOVINE, EYE, WHOLE, FRESH

8600822 Lampire Biologicals Laboratories 1EA
EUR 30.49

BOVINE, EYE, WHOLE, FROZEN

8620822 Lampire Biologicals Laboratories 1EA
EUR 30.49

Tissue cDNA, First Strand, Human Adult Normal, Adrenal, BioGenomics

MBS651970-40Tests MyBiosource 40Tests
EUR 680

Tissue cDNA, First Strand, Human Adult Normal, Adrenal, BioGenomics

MBS651970-5x40Tests MyBiosource 5x40Tests
EUR 2835

Pig Eye, Whole cDNA

PD-106 Zyagen 30 reactions
EUR 243

Sheep Eye, whole cDNA

SD-106 Zyagen 30 reactions
EUR 243

Rat Heart, whole cDNA

RD-801 Zyagen 30 reactions
EUR 243

Bovine Eye, Whole Total RNA

BR-106 Zyagen 0.1mg
EUR 160

MiniPig Eye, Whole cDNA

ND-106 Zyagen 30 reactions
EUR 358

Rat Stomach, whole cDNA

RD-302 Zyagen 30 reactions
EUR 243

Equine Brain, whole cDNA

ED-201 Zyagen 30 reactions
EUR 319

Rabbit Brain, whole cDNA

TD-201 Zyagen 30 reactions
EUR 243

Chicken Brain, Whole cDNA

CD-201 Zyagen 30 reactions
EUR 243

Rat Intestine, whole cDNA

RD-306 Zyagen 30 reactions
EUR 243

Rat Epididymis, whole cDNA

RD-402 Zyagen 30 reactions
EUR 243

Bovine Eye, Whole Total Protein

BT-106 Zyagen 1mg
EUR 153

Tissue cDNA, First Strand, Monkey (Rhesus) Adult Normal, Adrenal, BioGenomics

MBS652504-40Tests MyBiosource 40Tests
EUR 850

Tissue cDNA, First Strand, Monkey (Rhesus) Adult Normal, Adrenal, BioGenomics

MBS652504-5x40Tests MyBiosource 5x40Tests
EUR 3600

Immunoglobulin M (IgM) Whole Bovine

GWB-97A00C GenWay Biotech 1 mg Ask for price

Tissue cDNA, First Strand, Monkey (Cynomolgus) Adult Normal, Adrenal, BioGenomics

MBS652357-40Tests MyBiosource 40Tests
EUR 850

Tissue cDNA, First Strand, Monkey (Cynomolgus) Adult Normal, Adrenal, BioGenomics

MBS652357-5x40Tests MyBiosource 5x40Tests
EUR 3600

Bovine Adrenaline ELISA Kit

QY-E60146 Qayee Biotechnology 96T
EUR 511.2

Adrenal gland disease spectrum (adrenal gland cancer progression) tissue array (2017 WHO classification)

AD2081a TissueArray each
EUR 546
Description: Adrenal gland disease spectrum (adrenal gland cancer progression) tissue array (2017 WHO classification), including TNM and clinical stage (AJCC 7th edition), 96 cases/192 cores, replacing AD2081

Mouse CD1 Brain, whole cDNA

MD-201 Zyagen 30 reactions
EUR 243

Mouse C57 Brain, whole cDNA

MD-201-C57-1 Zyagen 30 reactions
EUR 280

Mouse CD1 Heart, whole cDNA

MD-801 Zyagen 30 reactions
EUR 243

Mouse C57 Heart, whole cDNA

MD-801-C57 Zyagen 30 reactions
EUR 280

Rat Spinal cord, whole cDNA

RD-230 Zyagen 30 reactions
EUR 243

Guinea Pig Brain, whole cDNA

GD-201 Zyagen 30 reactions
EUR 243

Mouse CD1 Stomach, whole cDNA

MD-302 Zyagen 30 reactions
EUR 243

Mouse BLC Stomach, whole cDNA

MD-302-BLC Zyagen 30 reactions
EUR 280

Mouse C57 Stomach, whole cDNA

MD-302-C57 Zyagen 30 reactions
EUR 280

Mouse CD1 Intestine, whole cDNA

MD-306 Zyagen 30 reactions
EUR 243

Mouse BLC Intestine, whole cDNA

MD-306-BLC Zyagen 30 reactions
EUR 280

Mouse C57 Intestine, whole cDNA

MD-306-C57 Zyagen 30 reactions
EUR 280

Mouse CD1 Epididymis, whole cDNA

MD-402 Zyagen 30 reactions
EUR 243

Mouse BLC Epididymis, whole cDNA

MD-402-BLC Zyagen 30 reactions
EUR 280

Mouse C57 Epididymis, whole cDNA

MD-402-C57 Zyagen 30 reactions
EUR 280

Adrenal tumors with normal adrenal tissue and matched cancer adjacent adrenal tissue array

AG991 TissueArray each
EUR 356
Description: Adrenal tumors with normal adrenal tissue and matched cancer adjacent adrenal tissue array, including TNM and clinical stage (AJCC 8th edition), 93 cases/99 cores (core size 1.5mm)

Adrenal Dissociation System 9 (Adrenal), Mouse and Rat

4-20209 CHI Scientific ea
EUR 195.92

Adrenal Lysate

21-160 ProSci 0.1 mg
EUR 468.6
Description: Monkey (Cynomolgus) adrenal tissue lysate was prepared by homogenization using a proprietary technique. The tissue was frozen in liquid nitrogen immediately after excision and then stored at -70°C. The monkey (Cynomolgus) adrenal tissue total protein is provided in a buffer including HEPES (pH 7.9), MgCl2, KCl, EDTA, Sucrose, Glycerol, Sodium deoxycholate, NP-40, and a cocktail of protease inhibitors. For quality control purposes, the adrenal tissue pattern on SDS-PAGE gel is shown to be consistent for each lot by visualization with coomassie blue staining. The adrenal tissue is then Western analyzed by either GAPDH or β-actin antibody, and the expression level is consistent with each lot.

Adrenal Lysate

1470 ProSci 0.1 mg
EUR 229.2
Description: Adrenal tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.

Adrenal Lysate

1417 ProSci 0.1 mg
EUR 229.2
Description: Adrenal tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.

Adrenal Lysate

MBS151591-01mg MyBiosource 0.1mg
EUR 200

Adrenal Lysate

MBS151591-5x01mg MyBiosource 5x0.1mg
EUR 880

Adrenal Lysate

MBS151750-01mg MyBiosource 0.1mg
EUR 200

Adrenal Lysate

MBS151750-5x01mg MyBiosource 5x0.1mg
EUR 880

Adrenal Lysate

XBL-11050 ProSci 0.1 mg
EUR 663.9
Description: Human adrenal tissue lysate was prepared by homogenization using a proprietary technique. The tissue was frozen in liquid nitrogen immediately after excision and then stored at -70°C. The human adrenal tissue total protein is provided in a buffer including HEPES (pH7.9), MgCl2, KCl, EDTA, Sucrose, Glycerol, Sodium deoxycholate, NP-40, and a cocktail of protease inhibitors. For quality control purposes, the adrenal tissue pattern on SDS-PAGE gel is shown to be consistent for each lot by visualization with coomassie blue staining. The adrenal tissue is then Western analyzed by either GAPDH or β-actin antibody, and the expression level is consistent with each lot.

Mouse CD1 Spinal cord, whole cDNA

MD-230 Zyagen 30 reactions
EUR 243

Mouse BLC Spinal cord, whole cDNA

MD-230-BLC Zyagen 30 reactions
EUR 280

Mouse C57 Spinal cord, whole cDNA

MD-230-C57 Zyagen 30 reactions
EUR 280

Bovine IgG Whole Molecule-Biotin Conjugate

MBS689526-1mg MyBiosource 1mg
EUR 185

Bovine IgG Whole Molecule-Biotin Conjugate

MBS689526-5x1mg MyBiosource 5x1mg
EUR 670

Rat Brain, whole Tissue cDNA

RD-201 Zyagen 30 reactions
EUR 243

Rat Brain, whole cDNA-Random Primer

RD-201-RH Zyagen 30 reactions
EUR 243

Rat Heart, whole cDNA-Random Primer

RD-801-RH Zyagen 30 reactions
EUR 243

Rat Stomach, whole cDNA-Random Primer

RD-302-RH Zyagen 30 reactions
EUR 243

Adrenal Dissociation System 2 (Leydig, Adrenal), Mouse and Rat

4-20202 CHI Scientific ea
EUR 161.87

Innovative Grade US Origin Bovine Whole Blood

IGBOWBK2E1000ML Innovative research each
EUR 312
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBK2E100ML Innovative research each
EUR 171
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBK2E500ML Innovative research each
EUR 230
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBK2E50ML Innovative research each
EUR 136
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBK3E1000ML Innovative research each
EUR 312
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBK3E100ML Innovative research each
EUR 171
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBK3E500ML Innovative research each
EUR 230
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBK3E50ML Innovative research each
EUR 136
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBKOF1000ML Innovative research each
EUR 312
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBKOF100ML Innovative research each
EUR 171
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBKOF500ML Innovative research each
EUR 230
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBKOF50ML Innovative research each
EUR 136
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBKOX1000ML Innovative research each
EUR 312
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBKOX100ML Innovative research each
EUR 171
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBKOX500ML Innovative research each
EUR 230
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBKOX50ML Innovative research each
EUR 136
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBLIH1000ML Innovative research each
EUR 312
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBLIH100ML Innovative research each
EUR 171
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBLIH500ML Innovative research each
EUR 230
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBLIH50ML Innovative research each
EUR 136
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBNAC1000ML Innovative research each
EUR 312
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBNAC100ML Innovative research each
EUR 171
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBNAC500ML Innovative research each
EUR 230
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBNAC50ML Innovative research each
EUR 136
Description: Innovative Grade US Origin Bovine Whole Blood

Innovative Grade US Origin Bovine Whole Blood

IGBOWBNAE1000ML Innovative research each
EUR 312
Description: Innovative Grade US Origin Bovine Whole Blood
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A new mathematical model for relative quantification in real-time RT-PCR.

Use of the real-time polymerase chain reaction (PCR) to amplify cDNA products reverse transcribed from mRNA is on the way to becoming a routine tool in molecular biology to study low abundance gene expression. Real-time PCR is easy to perform, provides the necessary accuracy and produces reliable as well as rapid quantification results. But accurate quantification of nucleic acids requires a reproducible methodology and an adequate mathematical model for data analysis.
This study enters into the particular topics of the relative quantification in real-time RT-PCR of a target gene transcript in comparison to a reference gene transcript. Therefore, a new mathematical model is presented. The relative expression ratio is calculated only from the real-time PCR efficiencies and the crossing point deviation of an unknown sample versus a control. This model needs no calibration curve. Control levels were included in the model to standardise each reaction run with respect to RNA integrity, sample loading and inter-PCR variations. High accuracy and reproducibility (<2.5% variation) were reached in LightCycler PCR using the established mathematical model.

One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products.

We have developed a simple and highly efficient method to disrupt chromosomal genes in Escherichia coli in which PCR primers provide the homology to the targeted gene(s). In this procedure, recombination requires the phage lambda Red recombinase, which is synthesized under the control of an inducible promoter on an easily curable, low copy number plasmid. To demonstrate the utility of this approach, we generated PCR products by using primers with 36- to 50-nt extensions that are homologous to regions adjacent to the gene to be inactivated and template plasmids carrying antibiotic resistance genes that are flanked by FRT (FLP recognition target) sites. By using the respective PCR products, we made 13 different disruptions of chromosomal genes.
Mutants of the arcB, cyaA, lacZYA, ompR-envZ, phnR, pstB, pstCA, pstS, pstSCAB-phoU, recA, and torSTRCAD genes or operons were isolated as antibiotic-resistant colonies after the introduction into bacteria carrying a Red expression plasmid of synthetic (PCR-generated) DNA. The resistance genes were then eliminated by using a helper plasmid encoding the FLP recombinase which is also easily curable. This procedure should be widely useful, especially in genome analysis of E. coli and other bacteria because the procedure can be done in wild-type cells.

Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes.

BACKGROUND
Gene-expression analysis is increasingly important in biological research, with real-time reverse transcription PCR (RT-PCR) becoming the method of choice for high-throughput and accurate expression profiling of selected genes. Given the increased sensitivity, reproducibility and large dynamic range of this methodology, the requirements for a proper internal control gene for normalization have become increasingly stringent. Although housekeeping gene expression has been reported to vary considerably, no systematic survey has properly determined the errors related to the common practice of using only one control gene, nor presented an adequate way of working around this problem.
RESULTS
We outline a robust and innovative strategy to identify the most stably expressed control genes in a given set of tissues, and to determine the minimum number of genes required to calculate a reliable normalization factor. We have evaluated ten housekeeping genes from different abundance and functional classes in various human tissues, and demonstrated that the conventional use of a single gene for normalization leads to relatively large errors in a significant proportion of samples tested. The geometric mean of multiple carefully selected housekeeping genes was validated as an accurate normalization factor by analyzing publicly available microarray data.

Analyzing real-time PCR data by the comparative C(T) method.

Two different methods of presenting quantitative gene expression exist: absolute and relative quantification. Absolute quantification calculates the copy number of the gene usually by relating the PCR signal to a standard curve. Relative gene expression presents the data of the gene of interest relative to some calibrator or internal control gene.
A widely used method to present relative gene expression is the comparative C(T) method also referred to as the 2 (-DeltaDeltaC(T)) method. This protocol provides an overview of the comparative C(T) method for quantitative gene expression studies. Also presented here are various examples to present quantitative gene expression data using this method.

 

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