SYBR Gold: Difference between revisions
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==General information== | ==General information== | ||
* | *ten-fold more sensitive than ethidium bromide. | ||
* | *greater dynamic range. | ||
* | *penetrates gels quickly. | ||
*Stains both DNA and RNA in conventional neutral polyacrylamide and agarose gels and in denaturing gels using urea, glyoxal or formaldehyde. | *Stains both DNA and RNA in conventional neutral polyacrylamide and agarose gels and in denaturing gels using urea, glyoxal or formaldehyde. | ||
* | *stain gels post-electrophoresis because SYBR Gold likely binds the charge phosphate backbone of nucleic acids altering the electrophoretic mobility significantly. (DNA bands can come out curved). | ||
* | *low background so no destaining is necessary. | ||
* | *can be excited at by standard transillumination at 300nm. | ||
* | *stained nucleic acids can be transferred to membranes for Northerns or Southerns. | ||
* | *doesn't inhibit most enzymatic reactions except PCR. PCR is sensitive to high dye concentrations. Address this issues by adjusting Mg<sup>2+</sup> concentration or remove stain via ethanol precipitation. | ||
* | *very expensive | ||
* | *supplied at 10,000X concentration in anhydrous DMSO. | ||
*operates most efficiently between pH 7-8.5 | |||
*stock solution is stable 6 months to 1 year at <= 20°C | |||
==References== | ==References== | ||
#[[Molecular Cloning]] | #[[doi:10.1101/pdb.prot4022|Molecular Cloning]] | ||
#[http://probes.invitrogen.com/media/publications/120.pdf Technical information from Invitrogen] | |||
#[http://probes.invitrogen.com/media/pis/mp11494.pdf SYBR Gold Nucleic Acid Gel Stain product info from Invitrogen] | |||
#[http://probes.invitrogen.com/media/pis/td004.pdf General SYBR stain tips and tricks from Invitrogen] | |||
Latest revision as of 15:12, 5 December 2006
Recommended by Molecular Cloning as the best of the SYBR dyes from Molecular Probes.
General information
- ten-fold more sensitive than ethidium bromide.
- greater dynamic range.
- penetrates gels quickly.
- Stains both DNA and RNA in conventional neutral polyacrylamide and agarose gels and in denaturing gels using urea, glyoxal or formaldehyde.
- stain gels post-electrophoresis because SYBR Gold likely binds the charge phosphate backbone of nucleic acids altering the electrophoretic mobility significantly. (DNA bands can come out curved).
- low background so no destaining is necessary.
- can be excited at by standard transillumination at 300nm.
- stained nucleic acids can be transferred to membranes for Northerns or Southerns.
- doesn't inhibit most enzymatic reactions except PCR. PCR is sensitive to high dye concentrations. Address this issues by adjusting Mg2+ concentration or remove stain via ethanol precipitation.
- very expensive
- supplied at 10,000X concentration in anhydrous DMSO.
- operates most efficiently between pH 7-8.5
- stock solution is stable 6 months to 1 year at <= 20°C
