単位の定義 SuperScript™ IV RTの1単位は、ポリ(A)オリゴ(dT)12-18をテンプレート/プライマーとして使用して、37℃で10分間に1 nmoleのデオキシリボヌクレオチドを酸沈殿性物質に組み込むために必要な酵素量です。
ユニットの反応条件 50 mM Tris-HCl(pH 8.3)、4 mM MgCl2、10 mM DTT、50 mM KCl、0.5 mM dTTP、0.4 MBq/mL [3H]-dTTP、0.4 mMポリ(A)オリゴ(dT)12-18および37℃で10分間の20 μL中の酵素。
研究用にのみ使用できます。診断用には使用いただけません。
仕様
最終産物タイプFirst-Strand cDNA
フォーマットStand-alone Enzyme
反応数50反応
最適反応温度50℃
数量10,000 Units
反応形態Separate components
試薬タイプReverse Transcription
逆転写酵素SuperScript™ IV
リボヌクレアーゼH活性低減
出荷条件ドライアイス
サイズ(最終製品)12.3 kb以下
原料RNA
技術Reverse Transcription
濃度200 U/μL
GC-Rich PCR Performance高
反応速度10 min.
Unit SizeEach
組成および保存条件
SuperScript™ IV RT(200 U/ÂμLで合計10,000ユニット)、50 ÂμL 5X RTバッファー、1 mL 0.1 MのDTT、500 ÂμL
-5℃~-30℃で保管してください。
よくあるご質問(FAQ)
Can I use a DNA-RNA hybrid as a template for M-MLV Reverse Transcriptase (Cat. No. 28025013, 28025021)? Can other reverse transcriptases, such as SuperScript reverse transcriptase, be used in the same way?
Yes, you can use a DNA-RNA hybrid as a template for M-MLV Reverse Transcriptase.
We have not tested this for SuperScript reverse transcriptases, so we cannot guarantee it would also work with those products.
This article can be used as a reference for additional information.
In your SuperScript IV RT protocols, there is no ezDNase inactivation step. Will active ezDNase affect RNA or the RT reaction?
The Invitrogen ezDNase Enzyme is a novel DNase that is highly specific for double-stranded DNA. It has no activity on single-stranded DNA in RT reactions (primers or probes), or on RNA. The enzyme is also thermolabile—it is inactivated quickly at temperatures typical for the SuperScript IV RT reaction (e.g., 50°C). The additional inactivation step is therefore not required in RT-qPCR applications.
Which SuperScript IV RT format do you recommend for real-time PCR applications?
For RT-qPCR applications we recommend using the Invitrogen SuperScript IV VILO Master Mix (Cat. No. 11756050). The cDNA synthesis reaction setup with this master mix requires fewer pipetting steps and therefore reduces variation in the data. SuperScript IV RT, as a component of the master mix, offers the highest efficiency of cDNA synthesis step compared to competitors’ products.
Are there any significant changes in the SuperScript IV RT protocol compared to the SuperScript III RT protocol?
The only change is that the incubation time for the reverse transcription reaction has been reduced from 50 minutes to 10 minutes. All the other parameters and steps are the same.
Can I get comparable cDNA yield and length using the SuperScript IV RT 10-minute protocol as when using the 50-minute protocol for SuperScript III RT?
When compared with SuperScript III RT (and other manufacturers’ RTs) in a synthesis reaction for a 9 kb cDNA, SuperScript IV RT performed successful synthesis in just 10 minutes and did so with comparable (or improved) yield (as shown by gel band density).