Go Back   Molecular Biology Forum > Molecular Research Topics Forum > Molecular Biology Techniques > PCR - Polymerase Chain Reaction Forum > Real-Time PCR and Quantitative PCR Forum
Register Blogs FAQ Members List Calendar Science Groups New! Arcade Search Today's Posts Mark Forums Read

Real-Time PCR and Quantitative PCR Forum Real-Time PCR and Quantitative PCR Forum


real-time Videos
real-time-pcr-protocol-of-gm-soybean-dna
real-time-pcr-protocol-of-gm-soybean-dna Real-Time PCR Protocol of GM Soybean DNA. From the Youtube user tangodurden.


Angiotensinogen RT-PCR protocol

Real-Time PCR and Quantitative PCR Forum

Real-Time PCR and Quantitative PCR Forum



Register Molecular Biology Forums
Reply
 
LinkBack Thread Tools Display Modes
  #1 (permalink)  
Old 06-19-2006, 04:31 AM
Pipette Filler
Points: 1,337, Level: 21Points: 1,337, Level: 21Points: 1,337, Level: 21
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Jun 2006
Posts: 1
vivek pande RSS Feed
Default Re: Expected band for PCR smears..

Could u please tell me about the Angiotensinogen RT-PCR protocol
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Alt Today
Advertising
Google Adsense
 
This advertising will not be shown
in this way to registered members.
Register your free account today
and become a member on
Molecular Biology Forum
Standard Sponsored Links

  #2 (permalink)  
Old 06-19-2006, 06:49 PM
admin's Avatar
Administrator
Points: 12,037, Level: 75Points: 12,037, Level: 75Points: 12,037, Level: 75
Activity: 100%Activity: 100%Activity: 100%
 
Join Date: Nov 2005
Posts: 1,010
Blog Entries: 3
admin RSS Feed
Default Re: Angiotensinogen RT-PCR protocol

I found this RT-PCR protocol:

Reverse-transcription PCR (RT-PCR)
RT-PCR for Angiotensinogen

First-strand cDNA was generated from 2.5 µg denatured total RNA with 20 U avian myeloblastosis virus reverse transcriptase, 1 µg random hexamers and reagents from Promega Corp. (50 min at 42 C and 4 min at 72 C). PCR primers, specific for renin (23), ACE (24, 25), cathepsin D (26), and cathepsin G (27) cDNA, were designed to span at least one intron. PCR was performed in 1x PCR buffer with 0.4 mmol/L deoxynucleotide triphosphate (Boehringer Mannheim, Mannheim, Germany), 1 µmol/L each of sense and antisense primers (Table 1), 2.5 U Taq polymerase (Boehringer Mannheim), and 2.5 U Pfu (Stratagene, La Jolla, CA), in a final vol of 50 µL, using a GeneAmpPCR system 9600 (Perkin-Elmer Corp., Norwalk, CT). Cycle conditions are available on request. Control samples for detection of possible PCR contamination were included in all RT-PCR experiments and were routinely negative.

Primers
Primer Sequence (5' to 3') Nucleotide position

hu Ang F TCC ACC TCG TCA TCC ACA 122–139

hu Ang R GGC TCC CAG ATA GAG AGA 433–450


Human Adipose Tissue Expresses Angiotensinogen and Enzymes Required for Its Conversion to Angiotensin II1

Cecilia Karlsson, Kajsa Lindell, Malin Ottosson, Lars Sjöström, Björn Carlsson and Lena M. S. Carlsson
The Journal of Clinical Endocrinology & Metabolism Vol. 83, No. 11 3925-3929

Last edited by admin; 06-19-2006 at 07:12 PM.
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
  #3 (permalink)  
Old 06-19-2006, 06:50 PM
admin's Avatar
Administrator
Points: 12,037, Level: 75Points: 12,037, Level: 75Points: 12,037, Level: 75
Activity: 100%Activity: 100%Activity: 100%
 
Join Date: Nov 2005
Posts: 1,010
Blog Entries: 3
admin RSS Feed
Default Re: Angiotensinogen RT-PCR protocol

Do a Pubmed Search for Angiotensinogen.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed
I am not sure what model (human, or mouse etc) you are using.

Last edited by admin; 06-19-2006 at 07:12 PM.
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Reply

Tags
angiotensinogen , protocol , rtpcr

Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On
Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Agarose gel electrophoresis of PCR products - Method and Protocol domba PCR - Polymerase Chain Reaction Forum 4 05-23-2008 04:42 PM
Protocol for preperation of fungal membranes/cell walls kra Signalling Biology 0 02-11-2008 07:17 PM
Protocol to prepare fungal membranes, cell walls kra Protocols and Methods Forum 0 02-11-2008 07:16 PM
Flow Chamber Assay Protocol moleculardude Cell Biology and Cell Culture 0 11-29-2006 12:02 AM
Matrigel Invasion Assay Protocol moleculardude Cell Biology and Cell Culture 0 11-28-2006 11:43 PM


All times are GMT. The time now is 11:50 PM.


Powered by vBulletin® Version 3.7.1
Copyright ©2000 - 2008, Jelsoft Enterprises Ltd.
Copyright 2005-2007 Molecular Station | All Rights Reserved