{"id":54787,"date":"2026-04-28T19:46:50","date_gmt":"2026-04-28T11:46:50","guid":{"rendered":"https:\/\/biopioneer.com.tw\/?p=54787"},"modified":"2026-04-28T19:46:50","modified_gmt":"2026-04-28T11:46:50","slug":"sybr-green-i-chloride-cas-2225748-05-8-mce%e8%b2%a8%e8%99%9fhy-d1191","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=54787","title":{"rendered":"SYBR green I chloride\u00a0 | CAS# 2225748-05-8 MCE\u8ca8\u865fHY-D1191"},"content":{"rendered":"

MCE (MedChem Express) Fluorescent Dye\u00a0<\/span> -SYBR green I chloride
\n<\/span><\/span><\/strong><\/span><\/p>\n

MCE (MedChemexpress) \u7e3d\u90e8\u4f4d\u65bc\u7f8e\u570b\u65b0\u6fa4\u897f\uff0c\u63d0\u4f9b\u7a2e\u985e\u9f4a\u5168\u7684\u5316\u5b78\u53ca\u751f\u5316\u7814\u7a76\u7528\u7522\u54c1\uff0c\u5305\u62ec\u5275\u65b0\u7684\u751f\u547d\u79d1\u5b78\u8a66\u5291\uff0creference compounds (\u6a19\u6e96\u5316\u5408\u7269)\uff0cAPIs (\u539f\u6599\u85e5) \u53caNatural compounds (\u5929\u7136\u5316\u5408\u7269)\u4ee5\u63d0\u4f9b\u5be6\u9a57\u5ba4\u6216\u79d1\u5b78\u7814\u7a76\u4e4b\u7528\uff0cMedChemExpress(MCE)\u7522\u54c150000+ \u7279\u7570\u6027\u6291\u5236\u5291\u3001\u5316\u5408\u7269\u5eab\u3001\u91cd\u7d44\u86cb\u767d\uff0c\u5c08\u6ce8\u65bc\u4fe1\u865f\u901a\u8def\u548c\u75be\u75c5\u7814\u7a76\u9818\u57df\u3002\u5168\u7403\u6587\u737b\u5ee3\u6cdb\u5f15\u7528\uff0c\u8cea\u63a7\u9ad4\u7cfb\u56b4\u683c\uff0c\u54c1\u8cea100%\u3002\u5b98\u7db2\u9023\u7d50<\/span> https:\/\/www.medchemexpress.com<\/a><\/span><\/p>\n

<\/p>\n

SYBR green I chloride | CAS# 2225748-05-8 MCE\u8ca8\u865fHY-D1191 <\/a><\/span><\/strong><\/span><\/p>\n

SYBR Green I chloride is a highly sensitive fluorescent nucleic acid dye that binds specifically to the minor groove of double-stranded DNA or intercalates between base pairs.<\/span><\/p>\n

\u7522\u54c1\u4ecb\u7d39: MCE\u54c1\u724c\u63d0\u4f9b\u7684\u4e00\u7cfb\u5217\u5316\u5408\u7269\uff0cSYBR Green I \u6c2f\u5316\u7269 (SYBR Green I chloride) \u662f\u4e00\u7a2e\u9ad8\u9748\u654f\u5ea6\u7684\u87a2\u5149\u6838\u9178\u67d3\u6599 (fluorescent nucleic acid dye)\uff0c\u53ef\u7279\u7570\u6027\u7d50\u5408\u96d9\u80a1 DNA \u7684\u5c0f\u6e9d\u6216\u5d4c\u5165\u9e7c\u57fa\u5c0d\u4e4b\u9593\u3002 SYBR Green I \u6c2f\u5316\u7269\u5728\u672a\u7d50\u5408\u72c0\u614b\u4e0b\u87a2\u5149\u8f03\u5f31\uff0c\u7d50\u5408\u5f8c\u5247\u767c\u51fa\u660e\u4eae\u7684\u87a2\u5149\uff0c\u4e26\u4e14\u512a\u5148\u7d50\u5408\u5927\u7247\u6bb5 DNA \u548c\u9ad8 G+C \u542b\u91cf\u7684 DNA\u3002 SYBR Green I \u6c2f\u5316\u7269\u9069\u7528\u65bc\u5373\u6642\u87a2\u5149\u5b9a\u91cf PCR \u6280\u8853 (real-time PCR technology)\uff1b\u5176\u87a2\u5149\u5f37\u5ea6\u8207\u64f4\u589e\u7522\u7269\u7684\u6578\u91cf\u548c\u5927\u5c0f\u76f8\u95dc\uff0c\u7121\u9700\u984d\u5916\u7684\u5f8c\u8655\u7406\uff0c\u5373\u53ef\u900f\u904e\u7194\u89e3\u66f2\u7dda\u5206\u6790\u5be6\u73fe\u57fa\u56e0\u8868\u73fe\u7684\u7cbe\u78ba\u5b9a\u91cf\u548c\u64f4\u589e\u5b50\u7684\u5340\u5206\u3002 SYBR Green I \u6c2f\u5316\u7269\u5ee3\u6cdb\u61c9\u7528\u65bc\u81e8\u5e8a\u524d\u9ad4\u5916\u6838\u9178\u6aa2\u6e2c ( in vitro nucleic acid detection)\u3002<\/p>\n

For research use only. We do not sell to patients.<\/p>\n

<\/p>\n

SYBR green I chloride<\/em>\u00a0Chemical Structure<\/p>\n

CAS No. :\u00a02225748-05-8<\/a><\/p>\n

1. Sample and Nucleic Acid Preparation<\/b><\/p>\n

Cell treatment:
\nSplenic monocytes are isolated from mice and divided into a control group and a PMA-ionomycin stimulation group, followed by incubation at 37\u2103 with 5% CO2<\/sub>\u00a0for 5 h.
\nRNA extraction:
\nCells are lysed with TRIZOL to extract total RNA; genomic DNA is removed with DNase I, RNA integrity is verified via denaturing gel electrophoresis, and RNA is quantified at 260 nm.
\ncDNA synthesis:
\n2 \u03bcg of RNA is annealed with 1 \u03bcg of oligo-dT; a reverse transcription system is added, and cDNA is synthesized through the procedure of 25\u2103 for 5 min \u2192 37\u2103 for 60 min \u2192 70\u2103 for 10 min.<\/p>\n

2. Reaction System Preparation<\/b>\u00a0(50 \u03bcL total volume)<\/p>\n

The system contains 0.5\u00d7SYBR Green I (10000\u00d7 stock solution), PCR buffer (50 mM KCl, 20 mM Tris-HCl pH 8.3, 2.5 mM MgCl2<\/sub>, 0.2% glycerol, 0.2% DMSO), 0.2 mM forward and reverse primers, 0.2 mM dNTPs, 1.25 U AmpliTaq, and 5 \u03bcL of 1:5 diluted cDNA.
\nNo-template controls (NTC) are set up, and all samples are run in at least 2 replicates.<\/p>\n

3. Amplification Cycling Program<\/b><\/p>\n

Pre-denaturation is performed at 95\u2103 for 30 s; 35 cycles of amplification are carried out (95\u2103 for 30 s \u2192 60\u2103 for 30 s \u2192 72\u2103 for 1 min \u2192 85\u2103 for 5 s).
\nFluorescence collection:
\nFluorescence is collected at 85\u2103 during each cycle (a temperature higher than the Tm of primer dimers and lower than the Tm of specific amplicons).<\/p>\n

4. Melting Curve Analysis<\/b><\/p>\n

After amplification, the temperature is increased from 60\u2103 to 95\u2103 at a rate of 2\u2103\/min, and fluorescence is detected every 15 s.
\nTarget amplicons and non-specific signals from primer dimers are distinguished based on specific Tm peaks.<\/p>\n

5. Quantification and Specificity Verification<\/b>
\nData analysis:
\nThe baseline is set to cycles 1-5, with a threshold of 0.7; Ct values are read by software, absolute copy numbers are calculated using an external plasmid standard curve (109<\/sup>-104<\/sup>\u00a0copies); \u03b2-actin is used as an internal reference for normalization to calculate relative expression levels.
\nSpecificity verification:
\nA single target band is confirmed via 2%\u00a0agarose<\/a>\u00a0gel electrophoresis, which matches the specific peak in the melting curve.<\/p>\n

Multiplex\u00a0Real-Time PCR<\/a>\u00a0Based on\u00a0Bacterial<\/a>\u00a0Samples<\/b>[2]<\/a><\/sup><\/p>\n

1. Template Preparation<\/b>
\nStrain culture: *Vibrio cholerae* is inoculated onto Columbia blood agar and cultured aerobically at 35\u00b0C for 24 h; *Legionella pneumophila* is inoculated onto BCYE agar and cultured aerobically at 35\u00b0C for 3 days.
\nDNA extraction: For *Vibrio cholerae*, 2-3 colonies are resuspended in 200 \u03bcL Instagene, and DNA is prepared following the kit instructions; for *Legionella pneumophila*, a\u00a0bacterial<\/a>\u00a0suspension with a turbidity of 0.5 McFarland standard is prepared as the template.<\/p>\n

2. Reaction System Preparation<\/b>\u00a0(20 \u03bcL total volume)
\nHomemade SYBR Green I (SG) premix:
\n1\u00d7 PCR Buffer II, 4 mM MgCl2<\/sub>, 200 \u03bcM dNTP, primers (0.5 \u03bcM each of *Legionella* 16S rRNA primers, 0.1 \u03bcM each of mip primers; 0.5 \u03bcM each of singleplex mip primers), 1 U AmpliTaq Gold, SG at a final concentration of 0.2\u00d7 or 0.4\u00d7 (from 10000\u00d7 stock solution), and 4 \u03bcL template DNA.<\/p>\n

3. Amplification Cycling Program<\/b>
\nFor *Vibrio cholerae*: Pre-denaturation at 95\u00b0C for 10 min; 40 cycles of 94\u00b0C for 20 s \u2192 60\u00b0C for 45 s \u2192 72\u00b0C for 45 s.
\nFor *Legionella pneumophila*: Pre-denaturation at 95\u00b0C for 10 min; 30 cycles of 94\u00b0C for 10 s \u2192 50\u00b0C for 20 s \u2192 72\u00b0C for 25 s.
\nFluorescence collection: Performed at 72\u00b0C in the SYBR channel (excitation at 470 nm, detection at 585 nm) with gains set to 2 and 5.<\/p>\n

4. Melting Curve Analysis
\nAfter amplification, the temperature is increased from 75\u00b0C to 99\u00b0C at a rate of 1\u00b0C per 60 s.
\nSoftware analysis: RotorGene V4.6.94 is used with digital filtration set to none, and the Tm value of amplicons is read.<\/p>\n

5. Product Verification
\n1%\u00a0agarose<\/a>\u00a0gel electrophoresis (with Gelstar dye added) is performed, and the melting curve is compared with the electrophoresis bands to verify the amplification of multiplex products.<\/p>\n

6. Key Usage Points
\nIn multiplex PCR, an SG concentration of **\u22650.4\u00d7** is required for simultaneous detection of multiple amplicons; a concentration of 0.2\u00d7 only detects products with high GC% and large fragment sizes.<\/p>\n

\"\u7db2\u7ad9\u898f\u683c\u5c0f\u5716_\u5de5\u4f5c\u5340\u57df<\/a><\/h4>\n

\"\"\u4ee3\u8b1d\u9175\u7d20\/\u86cb\u767d\u9175\u7d20 (Metabolic Enzyme\/Protease)\u00a0<\/span><\/span><\/strong><\/span><\/h4>\n