RT-PCR (including RNA preparation)

[DOC]

RT/PCR (including RNA preparation) – 9/14/04

File Format: Microsoft Word - View as HTML
RT PCR:. Kit used Platinum qRT-PCR ThermoScript One-Step System by Invitrogen. The following conditions are good starting points. ...
www.burnham.org/labs/wolf/Protocols/Protocols/Molecular%20Biology/RNA_preparation_RTPCR.doc -

Microarray experiments and quantitative RT PCR

[DOC]

Microarray experiments and quantitative RT PCR

File Format: Microsoft Word - View as HTML
Quantitative RT-PCR was performed to validate data from GeneChip experiments and was performed using an OPTICONTM 2 Continuous Fluorescence Detection System ...
www.nature.com/bjc/journal/v94/n10/extref/6603135x1.doc -

RNA extraction and RT-PCR

[DOC]

RNA extraction and RT-PCR

File Format: Microsoft Word - View as HTML
RNA extraction and RT-PCR. Total RNA from HeLa cells was prepared using the RNeasy Mini Kit (Qiagen) according to the manufacture's instruction. ...
www.nature.com/mp/journal/vaop/ncurrent/extref/4002052x4.doc

Protocol for Real-Time RT-PCR

[DOC]

Protocol for Real-Time RT-PCR

File Format: Microsoft Word - View as HTML
Reverse Transcription is carried out with the SuperScript First-Strand Synthesis System for RT-PCR. The following procedure is based on Invitrogen's ...
pga.mgh.harvard.edu/primerbank/PCR_protocol_short.doc -

Protocol for Real-Time RT-PCR

[DOC]

Protocol for Real-Time RT-PCR

File Format: Microsoft Word - View as HTML
This protocol describes the detailed experimental procedure for real-time RT-PCR using SYBR Green I as mentioned in Xiaowei Wang and Brian Seed (2003) A PCR ...
pga.mgh.harvard.edu/primerbank/PCR_protocol.doc

Quantitation of hepatitis C virus (HCV) Genome by RT-PCR

[DOC]

SOP for Quantitation of hepatitis C virus (HCV) Genome by RT-PCR

File Format: Microsoft Word - View as HTML
2.1 The HCV genome in HCV-infected cells is quantitated by RT-PCR using, as template, total RNA extracted from the cells, with competitor DNA, ...
www.brc.riken.jp/lab/dna/rvd/SOP/HCV/SOP-HCV-001EM.doc

RT-PCR

[DOC]

RT-PCR

File Format: Microsoft Word - View as HTML
RT-PCR. Total cellular RNA is isolated by lysing the pellet of tissue, cells, larvae, ... Dnase treatment of RNA sample prior to RT-PCR. RNA in water 1-8μl ...
www.biochem.northwestern.edu/holmgren/protocols/RT-PCR.doc

PCR & RT-PCR

[PDF]

PCR & RT -PCR

File Format: PDF/Adobe Acrobat - View as HTML
Template RNA for RT-PCR has to be free of DNA contamination. ... Always perform a control RT-PCR reaction with a RNA template that has not been ...
www.fermentas.com/pdf/catalog2006/3PCR.pdf -

Sample Prep for Real-Time RT-PCR

[PDF]

Sample Prep for Real-Time RT-PCR

File Format: PDF/Adobe Acrobat - View as HTML
RT-PCR, Northern, and RPA. Only different thing was using Polytron for ... RT inhibition of real-time PCR. Reverse transcriptase (1/10th the amount in a ...
www.abrf.org/Other/ABRFMeetings/ABRF2002/RNAprimer.pdf -

Single Tube Real-Time RT-PCR Assay

[PDF]

A Single Tube Real-Time RT-PCR Assay For The Detection Of ...

File Format: PDF/Adobe Acrobat - View as HTML
were stored frozen 24-48 hours before testing by the real-time RT-PCR method. ... Real-time RT-PCR was performed on the ABI Prism® 7900 HT Sequence ...
www.aruplab.com/Research&Development/resources/Posters/poster.2004.tubeentero.pdf -

Normalization of real-time quantitative RT-PCR data

[PDF]

Accurate normalization of real-time quantitative RT-PCR data by ...

File Format: PDF/Adobe Acrobat - View as HTML
accurately subtract the baseline value in real-time RT-PCR. data analysis. ... range of real-time RT-PCR methods, the requirements for a ...
www.embl.de/courses/qPCR/2007/vandesompele-normal%20GB02.pdf -

TaqMan One-Step RT-PCR Master Mix Reagents Kit Protocol

[PDF]

TaqMan One-Step RT-PCR Master Mix Reagents Kit Protocol P/N 4310299 D

File Format: PDF/Adobe Acrobat
The TaqMan One-Step RT-PCR Reagents Kit can be used for real- time ... Figure 1-1 Schematic representation of RT-PCR using the TaqMan One-Step ...
docs.appliedbiosystems.com/pebiodocs/04310299.pdf

Real-time RT-PCR

[PDF]

Real-time RT-PCR

File Format: PDF/Adobe Acrobat - View as HTML
OneStep RT-PCR buffer does not contain any components classified as ... OneStep RT-PCR Enzyme Mix contains glycerol, which Chemwatch reports as possibly ...
www.rsbs.anu.edu.au/.../MolecularBiologyFacility/NewProtocols/RealtimeRT-PCR(JohnShaw'smethod).pdf -

SARS-CoV Specific RT-PCR Primers

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SARS-CoV Specific RT-PCR Primers

File Format: PDF/Adobe Acrobat - View as HTML
negative RT-PCR controls, containing standardized viral RNA extracts, and nuclease-free. water were included in each run. Amplified 6-FAM-labeled products ...
www.who.int/csr/sars/CDCprimers.pdf -

Human Metapneumovirus RT-PCR Test

[PDF]

Human Metapneumovirus RT-PCR Test Now Available at YNHH

File Format: PDF/Adobe Acrobat - View as HTML
NOTE: Submit a separate sample when requesting HMPV RT-PCR. ... It is anticipated that HMPV RT-PCR will be available for clinical diagnosis ...
www.med.yale.edu/labmed/virology/newsletters/Newsletter13_1.pdf

Quantitative Real-Time RT-PCR

[PDF]

Quantitative Real-Time RT-PCR

File Format: PDF/Adobe Acrobat - View as HTML
Quantification of gene expression by real-time RT-PCR has. become the gold standard to which ... transcriptase-polymerase chain reaction (RT-PCR) amplifies ...
www.nanodrop.com/techsupport/ND-1000-Quantitative-Real-Time-RT-PCR-Application-Note.pdf -

Detection of Bornavirus

[PDF]

Detection of Bornavirus with the QIAGEN OneStep RT-PCR Kit

File Format: PDF/Adobe Acrobat
B the QIAGEN OneStep RT-PCR Kit. The reactions each contained the indicated concentration of nonspecific tRNA. Following reverse transcription, PCR ...
www1.qiagen.com/literature/qiagennews/0201/1016749_QNews22001p04-6.pdf -

Multiplex Titration RT-PCR

[PDF]

Multiplex Titration RT-PCR: Rapid Determination of Gene Expression ...

levels with RT-PCR. The procedure is rapid and does not require extensive .... Titration RT-PCR. While the relative expression levels determined with this ...
www.springerlink.com/index/J49KX27402517G37.pdf -

Quantitative RT-PCR from Rat Brain

[PDF]

Quantitative RT-PCR from Rat Brain

File Format: PDF/Adobe Acrobat - View as HTML
Quantitative RT-PCR. from Rat Brain. Procedure:. Flash frozen mouse spinal cord was cut into 30 µm sections. 10 motor neurons in mouse spinal cord were ...
www.leica-microsystems.com/.../$FILE/Application_note_Quantitative_RT_PCR.pdf -

RT Profiler PCR Arrays

[PDF]

RT Profiler PCR Arrays: Sensitive, Specific, and Reproducible ...

File Format: PDF/Adobe Acrobat - View as HTML
The Microarray Quality Control Consortium (MAQC) project and the real-time RT-PCR community have been defining optimal performance ...
www.superarray.com/manuals/pcrarraywhitepaper.pdf

REAL TIME RT-PCR PROTOCOL

[PDF]

REAL TIME RT-PCR PROTOCOL 1. PURPOSE This protocol describes how ...

File Format: PDF/Adobe Acrobat - View as HTML
REAL TIME RT-PCR PROTOCOL. SOP #: M011. REVISION LEVEL: .2. PAGE: 2 of 5. 4.1.2 RNases are very difficult to kill and can seriously affect the run. ...
compbio.dfci.harvard.edu/docs/RT-PCR-Quant.pdf

Quantitative real-time RT-PCR

[PDF]

REVIEW Quantitative real-time RT-PCR – a perspective

made real-time RT-PCR the benchmark technology for the detection and/or comparison of RNA levels. ... the unreliability of quantitative RT-PCR data, and its ...
jme.endocrinology-journals.org/cgi/reprint/34/3/597.pdf?ck=nck -

One-Step RT-PCR

[PDF]

One-Step RT-PCR

File Format: PDF/Adobe Acrobat - View as HTML
Prepare working primer stock solution as described in the two-step RT-PCR. protocol. There should be two negative controls among the PCRs. ...
ocw.mit.edu/NR/rdonlyres/Biology/7-16Spring-2005/302BAF49-4DC5-4492-8A07-1B867928730F/0/rt_pcr_1step.pdf

RT-PCR: Two-Step Protocol

RT-PCR: Two-Step Protocol Two-step Protocol

File Format: PDF/Adobe Acrobat - View as HTML
We will provide both one-step and two-step protocols for RT-PCR. ... In the one-step protocol, the components of RT and PCR are mixed in a single tube at ...
ocw.mit.edu/NR/rdonlyres/Biology/7-16Spring-2005/85278F14-478B-469E-8D2E-57E67E987F10/0/rt_pcr_2step.pdf -

Rapid RT-PCR

[PDF]

Rapid RT-PCR amplification from limited cell numbers.

phase were used for direct RT-PCR. Purified cells and colonies were lysed ... RT-PCR or frozen at -70. for later. use. By this method we were able to ...
www.genome.org/cgi/reprint/3/6/317.pdf

AgPathID One Step RT PCR

[PDF]

AgPathID One Step RT PCR.fm

File Format: PDF/Adobe Acrobat - View as HTML
The AgPath-ID™ One-Step RT-PCR Kit includes an enzyme. mixture, buffer, and Detection Enhancer ... RT-PCR can be used with the kit. You may need to optimize ...
www.ambion.com/techlib/prot/bp_1005.pdf -

Real-Time Reverse Transcriptase PCR Assays

[PDF]

Two Novel Real-Time Reverse Transcriptase PCR Assays for Rapid ...

Here we report the application of novel real-time RT-PCR assays for .... Real-time RT-PCR with Moloney murine leukemia virus RT and DNA Taq ...
jcm.asm.org/cgi/reprint/42/10/4759.pdf -

RT-PCR Kit

[PDF]

RT-PCR Kit

File Format: PDF/Adobe Acrobat - View as HTML
designed for high-fidelity RT-PCR. The kit contains the full ... was compared to the RT-PCR systems from two major suppliers. Total RNA ...
www.finnzymes.fi/pdf/phusion_rtpcrkit_prtpcr1_low.pdf

Amplifying RNA with RT-PCR

[PDF]

RNA Guide: Amplifying RNA with RT-PCR

File Format: PDF/Adobe Acrobat
RT-PCR amplification of human β-actin from mRNA isolated from human blood. mRNA was isolated from human blood using the PolyATtract ...
www.promega.com/guides/rna_guide/amplifyingrna.pdf

Real-time quantitative RT-PCR (Taqman)

[PDF]

Real-time quantitative RT-PCR (Taqman)

File Format: PDF/Adobe Acrobat - View as HTML
N.B. If more than one RNA sample is being used for RT-PCR, it is recommended. to prepare a master reagent mix. Some reactions may also be included ...
www.diabetesgenome.org/docs/Patti_Lab_Real-Time_qPCR_pr.pdf

RT PCR

[PDF]

RT PCR

File Format: PDF/Adobe Acrobat - View as HTML
RT PCR PROTOCOL: BD BIOSCIENCES. . This protocol is adapted from a BDBiosciences protocol by the Gene Expression Lab. This protocol is for use with BD ...
web.ncifcrf.gov/atp/LMT/RT_PCR_Protocol.pdf -

SYBR Green PCR Master Mix and RT-PCR

[PDF]

SYBR Green PCR Master Mix and RT-PCR

File Format: PDF/Adobe Acrobat - View as HTML
One-Step or Two-Step RT-PCR can be performed using the SYBR ...... nonspecific amplification in one-step RT-PCR using SYBR Green PCR ...
www.icmb.utexas.edu/core/DNA/Information_Sheets/Real-time%20PCR/7900syberProtocol.pdf

Real-Time PCR Vs. Traditional PCR

[PDF]

Real-Time PCR Vs. Traditional PCR

File Format: PDF/Adobe Acrobat - View as HTML
This tutorial will discuss the evolution of traditional PCR methods towards ... Advantages of Real-Time chemistries over traditional PCR. methods ...
www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdf

External RNA Quality Control by RT-PCR

[PDF]

External RNA Quality Control by RT-PCR

File Format: PDF/Adobe Acrobat - View as HTML
Control by RT-PCR. Walter Koch and Xiaoning Wu, Roche Molecular Systems ... RT-PCR using both 3' and 5' primer sets. (integrity) with and without RT step ...
www.cstl.nist.gov/div831/workshops/ERCC2003/10-QC.pdf

RT PCR and miniSTRs-McCord

PDF]

Microsoft PowerPoint - NEAFS Workshop RT PCR and miniSTRs-McCord.ppt

File Format: PDF/Adobe Acrobat - View as HTML Issues for quantitation by non RT-PCR. methods. • In spite of its use in mixture resolution,. PCR is not technically a quantitative. technique ... www.cstl.nist.gov/biotech/strbase/pub_pres/NEAFS_RTPCR_miniSTRs.pdf -

One-Step RT-PCR Kit

[PDF]

One-Step RT-PCR Kit

File Format: PDF/Adobe Acrobat - View as HTML
One-step RT-PCR is a variation on standard RT-PCR in which all components. of the RT and PCR are mixed in one tube prior to starting the reactions and in ...
www.usbweb.com/proto/78350a.pdf -

RT-PCR Methodology

RT-PCR Methodology

Animation to illustrate the way reverse transcriptase PCR (RT-PCR) produces DNA copies of an RNA template, from Cold Spring Harbor Laboratory Learning ...
www.bio.davidson.edu/Courses/immunology/Flash/RT_PCR.html - 6k - Cached

Reverse Transcriptase PCR Method

One-Step Reverse Transcriptase PCR Method for Detection of Borrelia burgdorferi mRNA in Mouse Lyme Arthritis Tissue Samples

F. X. Limbach,^1,2 B. Jaulhac,^1* Y. Piémont,¹ J. L. Kuntz,² H. Monteil,¹ and J. Sibilia²

Institut de Bactériologie de la Faculté de Médecine, Université Louis Pasteur et Hôpitaux Universitaires de Strasbourg,¹ and Service de Rhumatologie, Hôpitaux Universitaires de Strasbourg, ² 67000 Strasbourg, France

^*Corresponding author. Mailing address: Institut de Bactériologie, 3 rue Koeberlé, 67000 Strasbourg, France. Phone: 33.388.21.19.76. Fax: 33.388.25.11.13. E-mail: benoit.jaulhac@medecine.u-strasbg.fr .

 

http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=85022 

Optimized RT-PCR Method for Assaying Expression of Monocyte

Optimized RT-PCR Method for Assaying Expression of Monocyte Chemotactic Protein Type 1 (MCP-1) in Rabbit Aorta

Authors: Sękalska, B. ¹; Ciechanowicz, A.; Dołęgowska, B.; Naruszewicz, M.

Source: Biochemical Genetics, Volume 44, Numbers 3-4, April 2006 , pp. 129-139(11)

Publisher: Springer

http://www.ingentaconnect.com/content/klu/bigi/2006/00000044/F0020003/00009015

 

Protocol RT-PCR for detection and quantification of microRNAs

Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs

Erika Varkonyi-Gasic , Rongmei Wu , Marion Wood , Eric F Walton  and Roger P Hellens

Plant Methods 2007, 3:12doi:10.1186/1746-4811-3-12

Abstract

MicroRNAs (miRNAs) are a class of small non-coding RNAs with a critical role in development and environmental responses. Efficient and reliable detection of miRNAs is an essential step towards understanding their roles in specific cells and tissues. However, gel-based assays currently used to detect miRNAs are very limited in terms of throughput, sensitivity and specificity. Here we provide protocols for detection and quantification of miRNAs by RT-PCR. We describe an end-point and real-time looped RT-PCR procedure and demonstrate detection of miRNAs from as little as 20 pg of plant tissue total RNA and from total RNA isolated from as little as 0.1 ul of phloem sap. In addition, we have developed an alternative real-time PCR assay that can further improve specificity when detecting low abundant miRNAs. Using this assay, we have demonstrated that miRNAs are differentially expressed in the phloem sap and the surrounding vascular tissue. This method enables fast, sensitive and specific miRNA expression profiling and is suitable for facilitation of high-throughput detection and quantification of miRNA expression.

 

http://www.plantmethods.com/content/3/1/12

Real Time RT PCR

Why Real-Time RT-PCR?
Real-Time PCR Chemistries
Quantitation of Results
Instrumentation for Real-Time PCR
Tools for Real-Time RT-PCR
End-Point RT-PCR: Relative vs. Competitive vs. Comparative
Relative RT-PCR
Competitive RT-PCR
Comparative RT-PCR
Tools for Any RT-PCR Technique

 

Related Articles

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RNA Isolation for RT-PCR [read]

High Sensitivity qRT-PCR [read]

Designing a Successful qRT-PCR Experiment [read]

Top Ten Most Common Real-Time qRT-PCR Pitfalls [read]

 

ACETYLATION OF AMINO GROUPS ON PROTEINS

ACETYLATION (OR SUCCINYLATION) OF AMINO GROUPS ON PROTEINS

REFERENCE: Hanock and Benz. 1986. BBA. 860:699-707.
 

PURPOSE: Derivitization of amino groups to remove the positive charge characteristics of the protein, for example to determine the importance of amino groups to a protein's function.

http://www.cmdr.ubc.ca/bobh/methods/ACETYLATION.html