Quantification of nucleic acids: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
(comparison of RNA quant) |
||
| (One intermediate revision by the same user not shown) | |||
| Line 1: | Line 1: | ||
Nucleic acids are typically quantitated based on their absorbance of ultraviolet light. Each base has a different absorbance maximum, with the average being at 260 nm. Ambion has a good site that gives a variety of parameters for nucleotides and nucleic acids [http://www.ambion.com/techlib/append/na_mw_tables.html]. | Nucleic acids are typically quantitated based on their absorbance of ultraviolet light. Each base has a different absorbance maximum, with the average being at 260 nm. Ambion has a good site that gives a variety of parameters for nucleotides and nucleic acids [http://www.ambion.com/techlib/append/na_mw_tables.html]. | ||
== Quantification of RNA == | |||
* [[RNA blot]]ting or Northern blotting | |||
* [[ribonuclease protection assays]] (RPA) | |||
* [[Q-PCR]] or more specifically [[QRT-PCR]] | |||
RNA blotting and RPAs are the gold standards, since no amplification is involved, but require more RNA. Q-PCR is the most sensitive method and can discriminate closely related mRNAs. It is therefore suitable for quickly quantifying low amounts of RNA. | |||
== Quantification of DNA == | |||
* [[DNA blot]]ting or Southern blotting | |||
* [[Q-PCR]] | |||
Latest revision as of 10:15, 21 February 2007
Nucleic acids are typically quantitated based on their absorbance of ultraviolet light. Each base has a different absorbance maximum, with the average being at 260 nm. Ambion has a good site that gives a variety of parameters for nucleotides and nucleic acids [1].
Quantification of RNA
- RNA blotting or Northern blotting
- ribonuclease protection assays (RPA)
- Q-PCR or more specifically QRT-PCR
RNA blotting and RPAs are the gold standards, since no amplification is involved, but require more RNA. Q-PCR is the most sensitive method and can discriminate closely related mRNAs. It is therefore suitable for quickly quantifying low amounts of RNA.
Quantification of DNA
- DNA blotting or Southern blotting
- Q-PCR
