SYBR Green I Staining of DNA in Gels

2014/12/08 admin 88

SYBR Green I Gel Staining Solution, 10000x

 
简介:
SYBR Green I是一种与dsDNA结合具有极高灵敏度的染料,其可以用于琼脂糖凝胶和聚丙烯酰胺凝胶电泳的DNA检测。不同于乙锭类(Ethidium)染料的是,SYBR Green I为无毒的染料,与双链DNA(double stranded DNA)具有更高的结合力,因此表现出更高的灵敏度,
 
订购及保存信息:
货号
浓度
规格
保存条件:
HC0251
10000x
50ul / 250ul/1ml
-20C避光保存1年;可常温运输条件下3周有效。勿长时间暴露于光下。
 
技术参数:
外观:
橙色溶液
质控:
NMR 1H (95%) and 13C, TLC, functional testing
最大激发波长(nm):
454
最大激发波长时的消光系数(Lmol-1cm-1):
73000
最大发射波长(nm):
524
荧光量子产率:
0.8
 
 
溴化乙锭(Ethidium Bromide)和SYBR Green I的参数对比:
Feature
Ethidium Bromide
SYBR Green I
Fluorescence
Red (615 nm)
Green (524 nm)
Excitation maximum
302 nm
454 nm
Excitation light source
UV only
Blue light or UV
Sensitivity
2 ng / band(dsDNA)
100 ng / band (RNA)
0.08 ng / band (dsDNA)
1-2 ng / band (oligonucleotides)
Health hazard
High
Low
 
 
推荐实验手册:
Protocol: SYBR Green I Staining of DNA in Gels
 
SYBR Green I是一种与双链DNA特异性结合的荧光染料。有三种染色方案可以选择:凝胶浸泡法;凝胶预染色;样本预染色。
SYBR Green I is fluorescent dye that binds specifically to double-stranded DNA. There are three variants of staining protocol: gel soaking, gel pre-staining, and sample pre-staining.
 
Ð凝胶浸泡法;
ÐGel soaking:
 
琼脂糖凝胶电泳和聚丙烯酰胺凝胶电泳的经典方法:
Classical method for agarose and polyacrylamide gels.
 
1.在琼脂糖凝胶或聚丙烯酰胺凝胶中电泳样本。
Run sample(s) in agarose or polyacrylamide gel.
2.在烧杯中,取10,000x SYBR Green I溶液10ul加入100ml的1x TE,TBE或TAE缓冲液(mini凝胶用);或者取10,000x SYBR Green I溶液50ul加入500ml的1x TE,TBE或TAE缓冲液(中等大小的凝胶用)。使用小铲、玻璃棒或磁力搅拌器完全混匀。
In a beaker, add 10uL of 10,000x SYBR Green I solution in DMSO to 100 mL of 1x TE, TBE, or TAE buffer (for mini gel), or 50uL of 10,000x SYBR Green I solution in DMSO to 500 mL of 1x TE, TBE, or TAE buffer (for mid-sized gel). Mix thoroughly with spatula, rod, or magnetic stirrer.
3.将稀释好的SYBR Green I溶液倒入合适大小的托盘。
Pour the diluted SYBR Green I solution in appropriate tray or pan.
4.将凝胶浸泡在SYBR Green I溶液中5-10min。
Soak the gel for 5-10 min.
5.在254nm低压汞灯和橙色滤光片下观察或记录凝胶。
View or document the gel using 254 nm low-pressure mercury lamp and orange filter.
 
Ð凝胶预染色:
ÐGel pre-staining:
 
此方法仅用于琼脂糖凝胶电泳,不适用于PAAG.
This method is acceptable for agarose gels only, but not for PAAG.
 
1.使用微波炉或加热器煮沸缓冲液来溶解琼脂糖。
Boil the agarose in buffer to dissolution using microwave or heating appliance.
2.加热溶解后,每10ml凝胶溶液加入10,000x SYBR Green I溶液1ul。完全混匀。
3.While hot, add 1uL of 10,000x SYBR Green I solution in DMSO per each 10 mL of gel solution. Mix thoroughly.
4.灌胶并使其凉至室温。
Pour the gel and let it cool down.
5.为得到更好的实验结果,可以在正极(“+”,红线)附件每10ml缓冲液加入10,000x SYBR Green I溶液1ul。
For best results, add 1uL of 10,000x SYBR Green I solution in DMSO per each 10 mL of buffer near anode ("+", red wire).
6.跑胶。在254低压汞灯下实时监测可能的迁移带。
Run the samples. Real-time monitoring of migrating bands under 254 nm low-pressure mercury lamp possible.
7.在254nm低压汞灯和橙色滤光片下观察或记录凝胶。
View or document the gel using 254 nm low-pressure mercury lamp and orange filter.
 
 
Ð样本预染色:
ÐSample pre-staining:
 
灵敏度较低,最经济型的方法。
Least sensitive, most economical method.
 
1.混匀25ul DMSO和10,000x SYBR Green I溶液1ul。
Mix 25uL of DMSO and 1uL of 10,000x SYBR Green I solution in DMSO.
2.每个由琼脂糖凝胶或聚丙烯酰胺凝胶分离的样本中加入混匀好的该溶液1ul。
Add 1uL of the solution to each sample to be separated on agarose or polyacrylamide gel.
3.跑胶。在254低压汞灯下实时监测可能的迁移带。
Run the samples. Real-time monitoring of migrating bands under 254 nm low-pressure mercury lamp possible.
4.在254nm低压汞灯和橙色滤光片下观察或记录凝胶。
View or document the gel using 254 nm low-pressure mercury lamp and orange filter.
 
参考文献:
Note: fluorescent properties of SYBR Green I bound to dsDNA below are taken from publication:
Zipper, H.; Brunner, H.; Bernhagen, J.; Vitzthum, F. Investigations on DNA intercalation and surface binding by SYBR Green I, its structure determination and methodological implications. Nucleic Acids Res., 2004, 32, e103.
 
 
【备注:本文中文翻译仅供参考,以对照的英文说明为准。所有产品仅供科研应用。不用于临床诊断或治疗。】