
Sirius DNA Transfection Reagentis a multi-component reagent that forms a complex with DNA and siRNA , then transports the complex insect cells.
Storage:Store at 4°C (Don’t Freezing).
Functional analysis:Cos 7 cells are transfected in 96 well plates with a reporter gene vector DNA, using 0.2-0.5 μlSirius Transfection Reagent and 0.1 μg DNA or 0.25 μl
Sirius Transfection Reagentand 0.1 μg DNA. Reporter gene activity is monitored via a colorimetric test against a reference lot. Activity must be 70% - 90%
compared to a reference lot in at least one of the tested amounts, listed above.
ProcedureDetails | ||||
Component | 96-well | 24-well | 12-well | 6-well |
Adherentcells | 1–4×104 | 0.5–2×105 | 0.25–1×106 | |
Opti-MEM®Medium | 25μL×4 | 50μL×4 | 100μL×4 | 150μL×4 |
Sirius TransfectionReagent | 1,1.5,2,2.5μL | 2,3,4,5μL | 4,6,8,10ul | 6,9,12,15μL |
Opti-MEM®Medium | 125μL | 250μL | 500ul | 700μL |
DNA(0.5–5μg/μL) | 2.5μg | 5μg | 10ug | 14μg |
DilutedDNATotal | 25μL | 50μL | 100ul | 150μL |
DilutedSirius TransfectionReagent | 25μL | 50μL | 100ul | 150μL |
Incubatefor5minutesatroomtemperature. | ||||
Component | 96-well | 24-well | 12-well | 6-well |
DNA-lipidcomplexperwell | 10μL | 50μL | 100ul | 250μL |
FinalDNAusedperwell | 100ng | 500ng | 1000ng | 2500ng |
FinalSirius TransfectionReagentusedperwell | 0.2–0.5μL | 1.0–2.5μL | 2.0-5.0ul | 5.0–12.5μL |
Incubatecellsfor1–3daysat37°C.Thenanalyzetransfectedcells. |
analysis is tested for viability.
Sirius DNA Transfection Reagentcomplexes must be made in serum-free medium such as Opti-MEM Reduced serum Medium and can be added directly to cells in
culture medium, in the presence or absence of serum .
It is not necessary to remove complexes or change and add medium after transfection.The amount of Sirius Transfection Reagentrequired for successful
transfection varies depending on the cell type and passage number. Start any new transfection by testing the recommended four concentrations of Sirius DNA to
determine an optimum amount.
Sirius DNA Transfection Reagent is a proprietary formulation for transfectin nucleic aicids into a wide range of eukaryotic cells . It is designed to transfect a
wide range of eukaryotic cells including insect cells and many cell lines not transfected well by other reagents (e.g.,MCF-7, RAW 264.7, PC-3, HeLa, MA-10,
HepG2, SH-SY5Y, A7r5, STO, SCC-61, STSAR-90, SQ20B, T98).
Note: For the most up-to-date list of cell types that have been successfully transfected with Sirius DNA Transfection Reagent for life science research only.
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TAKARA的RR047A逆转录试剂盒,做逆转录程序设置错误本来应该37℃15min85℃5s,变成37℃15s,85℃
5s,请问再重新设置程序可以吗,85℃酶会全部失活吗?
加尾法是采用加A酶先对mirna进行加尾,然后再用带oligodt的引物反转录,他的反转录引物是通用的,一次反转录可以获得所有miRNA的cdna,效率高。茎环法是采用特异性反转录引物序列+颈环结构作为反转录引物进行反转录的,一次反转录只能获得一种mirna的cdna,可能有几种一起反转录的,这个我不太清楚一起反转录的效果。然后茎环法的经典即ABI的探针法定量试剂盒。我用过茎环法反转录+染料法定量的,下游引物为通用引物,结果不太好,就现在用的这家的他采用的是双向引物都是特异性的,效果还可以。不知道我说的清楚不,希望能对你有帮助。
向左转|向右转
逆转录(reverse transcription)是以RNA为模板合成DNA的过程,即RNA指导下的DNA合成。此过程中,核酸合成与转录(DNA到RNA)过程与遗传信息的流动方向(RNA到DNA)相反,故称为逆转录。逆转录过程是RNA病毒的复制形式之一,需逆转录酶的催化。 逆转录过程的揭示是分子生物学研究中的重大发现,是对中心法则的重要修正和补充。人们通过体外模拟该过程,以样本中提取的mRNA为模板,在逆转录酶的作用下,合成出互补的cDNA,构建cDNA文库,并从中筛选特异的目的基因。该方法已成为基因工程技术中最常用的获得目的基因的策略之一。
大家miRNA逆转录和qPCR的试剂盒用的是那个公司的啊,求推荐!

