Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Basic Lab Protocols and Techniques
Register Search Today's Posts Mark Forums Read

primer resuspension, buffers -step by step

primer resuspension, buffers -step by step - Basic Lab Protocols and Techniques

primer resuspension, buffers -step by step -

LinkBack Thread Tools Display Modes
Old 06-17-2011, 09:04 AM
Pipette Filler
Points: 69, Level: 1 Points: 69, Level: 1
Activity: 0% Activity: 0% Activity: 0%
Join Date: Jun 2011
Location: Budapest
Posts: 2
Thanks: 6
Thanked 0 Times in 0 Posts
Default primer resuspension, buffers -step by step

Hi All,

Could you please help me how to prepare TAE (50X) and TE buffer?

I have already found a recipe for TAE here among the threads, so is it the right one?:
"First you need to know preparation of 0.5M EDTA solution:
Add 186.1 g EDTA (disodium, dihydrate, ) to 800 ml of ddH20.
Add about 20g of NaOH pellets while stirring to bring the pH to 8.0.
Add the last few grams slowly to avoid overshooting the pH.
Note that the EDTA won't completely dissolve until the pH is around 8,
make the final volume to 1000ml with ddH2o.
Filter with 0.5 micron filter and autoclave

50 X TAE Buffer Preparation protocol (Tris-Acetate-EDTA)

242 gm - Tris base
57.1 mL - Acetic Acid
100mL - 0.5 M EDTA (shake vigorously before use)

Add ddH2O to 1 Liter and adjust ph to 8.5 using KOH."

I would also like to resuspend our oligos (primers) in TE according to the newletter of the company for long term storage I am to use : TE buffer (10mM Tris, 0,1mM EDTA pH 8.0.)

Since I was given the task to start the molbio lab without an experienced laboratory technician and without lab practice (just with theory backgorund: molecular biologist, got the diploma 10 years ago, and hs worked on different field until now) I need to spend endless time to try to clarify all small questions, and after spending days with reading I am more and more scared....

Could you pelase help me?

Thank you very much in advance,
Reply With Quote
Old 08-06-2011, 05:20 AM
Points: 231, Level: 4 Points: 231, Level: 4
Activity: 0% Activity: 0% Activity: 0%
Join Date: Aug 2011
Location: Malaysia
Posts: 36
Thanks: 0
Thanked 14 Times in 13 Posts
Default Re: primer resuspension, buffers -step by step

The 50x TAE buffer protocol is correct. EDTA can only dissolve when the pH reaches 8. Yes, for long term primer stock storage at -20C, it is better to dissolve the lyopholized primers in TE buffer. Then you can dilute you primers in sterile water for PCR work from the stock.
Reply With Quote

buffers , primer , resuspension , step , tae

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
Bad primer design ? How can I know ? surt Bioinformatics 4 10-07-2009 07:24 AM
Primer Dimers Lisa PCR - Polymerase Chain Reaction Forum 2 03-25-2009 09:10 PM

All times are GMT. The time now is 01:57 AM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2013, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.16511 seconds with 16 queries