KudoZ home » English » Science (general)

ambient buffer, stringent buffer

English translation: low-stringency buffer, high-stringency buffer

Login or register (free and only takes a few minutes) to participate in this question.

You will also have access to many other tools and opportunities designed for those who have language-related jobs
(or are passionate about them). Participation is free and the site has a strict confidentiality policy.
GLOSSARY ENTRY (DERIVED FROM QUESTION BELOW)
English term or phrase:ambient buffer, stringent buffer
English translation:low-stringency buffer, high-stringency buffer
Entered by: Jörgen Slet
Options:
- Contribute to this entry
- Include in personal glossary

12:13 Oct 22, 2004
English to English translations [PRO]
Medical - Science (general) / nucleic acid hybridisation
English term or phrase: ambient buffer, stringent buffer
"4. Prepare Working ***Stringent*** Wash Buffer as follows: Examine SSPE and SDS and, if necessary, warm to 50°C in a water bath to redissolve any precipitate. Add 25 mL of SSPE to 965 mL of distilled or deionized water. Mix well. Add 10 mL of SDS and mix well. The Working Stringent Wash Buffer is sufficient for 100 LINEAR ARRAY HCV Genotyping Strips. Working Stringent Wash Buffer should be stored at room temperature in a clean container and is stable for 30 days.

5. Prepare Working ***Ambient*** Wash Buffer as follows: Examine SSPE and SDS and, if necessary, warm to 50°C in a water bath to dissolve any precipitate. Add 37.5 mL of SSPE to 1447.5 mL of distilled or deionized water. Mix well. Add 15 mL of SDS and mix well. The Working Ambient Wash Buffer is sufficient for 100 LINEAR ARRAY HCV Genotyping Strips. Working Ambient Wash Buffer should be stored at room temperature in a clean container and is stable for 30 days."
______________

Could you explain the meanings of these two, especially the "ambient buffer" ?
Jörgen Slet
Estonia
Local time: 18:40
buffer salt concentration affects the 'effective melting temp.'.....
Explanation:
Hi Jörgen

Firstly, 'ambient' wash buffer is rarely used. Depending on the salt concentration, this would normally be called a low-stringency buffer. The stringency refers to the 'effective melting temperature' of a hybridized nucleic acid. Basically, the temperature at which a hybridized nucleic acid probe will 'melt' or come away from the target that it is bound to depends on two things: the similarity of the probe and its target, and the salt concentration of the buffer. At higher salt concentrations, the 'effective Tm' is higher, which means that when you wash off randomly bound 'probes', more will stay bound - the buffer is less stringent. At low salt concentrations, in a stringent buffer, the 'effective Tm' is more dependent upon the similarity between the probe and its target, so when you wash with this buffer, more non-specifically bound probe will be washed away.

Usually what happens is you gradually reduce the stringency of the washing buffers so that you end up with a specific signal, but without washing nearly all of it away.

Quite a lot of this is explained in this link:
http://www.ndsu.nodak.edu/instruct/mcclean/plsc731/dna/dna6....

I hope this helps.

All the best.
Selected response from:

xxx
Grading comment
Thanks, this was quite helpful !
4 KudoZ points were awarded for this answer



Summary of answers provided
5 +1buffer salt concentration affects the 'effective melting temp.'.....
xxx


Discussion entries: 2





  

Answers


2 hrs   confidence: Answerer confidence 5/5 peer agreement (net): +1
buffer salt concentration affects the 'effective melting temp.'.....


Explanation:
Hi Jörgen

Firstly, 'ambient' wash buffer is rarely used. Depending on the salt concentration, this would normally be called a low-stringency buffer. The stringency refers to the 'effective melting temperature' of a hybridized nucleic acid. Basically, the temperature at which a hybridized nucleic acid probe will 'melt' or come away from the target that it is bound to depends on two things: the similarity of the probe and its target, and the salt concentration of the buffer. At higher salt concentrations, the 'effective Tm' is higher, which means that when you wash off randomly bound 'probes', more will stay bound - the buffer is less stringent. At low salt concentrations, in a stringent buffer, the 'effective Tm' is more dependent upon the similarity between the probe and its target, so when you wash with this buffer, more non-specifically bound probe will be washed away.

Usually what happens is you gradually reduce the stringency of the washing buffers so that you end up with a specific signal, but without washing nearly all of it away.

Quite a lot of this is explained in this link:
http://www.ndsu.nodak.edu/instruct/mcclean/plsc731/dna/dna6....

I hope this helps.

All the best.

xxx
Specializes in field
Native speaker of: Native in EnglishEnglish
PRO pts in category: 8
Grading comment
Thanks, this was quite helpful !

Peer comments on this answer (and responses from the answerer)
agree  Nizamettin Yigit
6 hrs
Login to enter a peer comment (or grade)




Return to KudoZ list


KudoZ™ translation help
The KudoZ network provides a framework for translators and others to assist each other with translations or explanations of terms and short phrases.



See also:



Term search
  • All of ProZ.com
  • Term search
  • Jobs
  • Forums
  • Multiple search