產(chǎn)品名稱:DNA電泳試劑 SYBR Green II(10000×)
產(chǎn)品型號(hào):SY1040
產(chǎn)品特點(diǎn):DNA電泳試劑 SYBR Green II(10000×)SYBR ® Green II RNA gel stainCat# :SY1020Introduction SYBR
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DNA電泳試劑 SYBR Green II(10000×)
SYBR ® Green II RNA gel stain
Cat# :SY1020
Introduction
SYBR
® Green II RNA gel stain is one of the most sensitive dyes known for detecting RNA in electrophoreticgels. As little as 100 pg RNA or single-stranded DNA per band can be detected in a SYBR Green IIBstained agarose or polyacrylamide gel using 254 nm epi-illumination Polaroid
® 667, black-and-white print film and a SYBR Green gel stain photographic filter.
Even with 300 nm transillumination, as little as 500 pg RNA per band can be detected. SYBR Green II RNA gel stain is significantly more sensitive than ethidium bromide,the most commonly used stain for detecting nucleic acids in gels (Figure 1). With 300 nmtransilluminationand photography through an orange-red gelatin filter, ethidium bromide’s sensitivity limit in a standardagarose minigel is about 1.5 ng single-stranded nucleic acid per band. Note that our detection limits arebased on results obtained with a FotoDyne
® Foto/UV ® 450 ultraviolet transilluminator in combination with
Polaroid 667 film. Video cameras and CCD cameras in general have a different spectral response thanblack-and-white print film and thus may not exhibit the same sensitivity.
A BFigure 1. Dilution series of Escherichia coli ribosomal 16S and 23S RNA electrophoresed in 1% agarose gels. The gelscontain an identical twofold dilution series of RNA. The gel shown in panel A was stained for 20 minutes with SYBR Green II
RNA gel stain (using a 1:10,000 dilution of the stock reagent) and not destained. The gel shown in panel B was stained for 20minutes with 5 µg/mL ethidium bromide, then destained for an additional 20 minutes. The SYBR Green II dye-stained gel was excited using 254 nm epi-illumination and the ethidium bromide–stained gel using 300 nm transillumination (Fotodyne
® FotoUV 450 ultraviolet transilluminator). Although the SYBR Green II dye-stained gel can be excited at 300 nm, epi-illumination at 254 nm resulted in the best sensitivity in our hands. Both gels were photographed with Polaroid 667 black-and-white print film,using a SYBR Green gel stain photographic filter (SYBR Green II dye-stained gel) or an ethidium bromide gel stainphotographic filter (ethidium bromide– stained gel).
SYBR Green II RNA gel stain can be used to:
Analyze small aliquots from RNA preparations before Northern blotting, start-site mapping or cDNApreparation
Visualize the migration behavior of 5S rRNA species after high-resolution denaturing gradienectrophoresis (DGGE)
Stain DNA in single-strand conformation polymorphism (SSCP)
Stain DNA before amplification by PCR
Materials Contents SYBR Green II stain is provided as a 10,000X concentrate in DMSO in the following sizes:
50 µL (SY1040-50)
500 µL (SY1040-500)
Storage
Upon receipt, store the dye frozen at -20°C, protected from light in a desiccator. When stored properly,the SYBR Green II stain stock solution in DMSO is stable for six months to one year.
注:SYBR Green II主要用于RNA和單鏈DNA電泳。是EB和替代品,無致癌性,超敏感性。用作常規(guī)電泳時(shí),須將商品化的核酸MARKER作一定倍數(shù)的稀釋(1/5-1/10),這樣才能得到比較理想的結(jié)果。
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