SYBR green problem.. - real time PCR SYBR (Aug/11/2006 )

hi everyone

How are you use SYBR green concentration in home-made mix?

i use 1X....but interruption...

and If use 0.3X, I got a product but do not detect fluorescence or less fluorescence

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SYBR Green I Nucleic
Acid Gel Staina
! Supplieda as a stock solution in DMSO, stable at – 20°C
for 6 – 12 months.
! Working solution:
dilute the stock solution 1:10,000 in TE, TBE or TAE buffer.
! Sensitivity:
as low as 80 pg per band dsDNA using 312 nm transillumination with the
Lumi-Imager F1 System*.

* roche genetics


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SYBR Green II RNA
Gel Staina
! Supplieda as a stock solution in DMSO stable at – 20°C
for 6 – 12 months.
! Working solution:
For nondenaturating gels and denaturing polyacrylamide/urea gels:
1:10,000 dilution in TBE
For denaturing agarose/formaldehyde gels:
1:5,000 dilution in TBE
! Sensitivity:
as low as 5 ng per band of RNA using 312 nm transillumination with the
Lumi-Imager F1 System**

** roche genetics

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There are some latest ideas to replace SYBR by SYTO9 in real time
SYTO9 is "saturating" and less inhibiting

see NAR paper and related

in Corbett for RotorGene-6000 a report for SYTO9

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