【RScript cDNA 合成試劑盒(200 U/μl) 】 RScript cDNA Synthesis Kit貨號RK001-0050
RScript cDNA 合成試劑盒(200 U/μl) | RScript cDNA Synthesis Kit貨號RK001-0050
RScript cDNA Synthesis Kit (50 Rxns)
Categories: RNA Reagent
一、產品介紹
RScript cDNA 合成試劑盒(RScript cDNA Synthesis Kit, RK001-0050) 專門用於研究和診斷應用，可滿足所有的 cDNA 合成需求；克服寬溫度範圍內最具挑戰性的二級 RNA 結構(secondary RNA structures)。套組內包含新一代工程重組 M-MLV 逆轉錄酶(M-MLV reverse transcriptase)，具有改進的熱穩定性( improved thermostability)、持續合成能力(robustness)、穩健性、最佳 cDNA 產量( optimal cDNA yields)、Moloney 小鼠白血病病毒逆轉錄酶 (M-MLV RT) 缺乏 DNA 核酸內切酶活性，具有較低的 RNase H 活性。是最通用的世界上的逆轉錄酶，不僅可以簡單地滿足常規 cDNA 合成要求，而且即使對於手頭最具挑戰性的 RNA 樣品也能實現卓越的性能。
Reverse transcription of a 330 bp fragment CCD1A RNA using RScript cDNA Synthesis Kit to carry this experiment. The PCR data shows that RScript cDNA Synthesis Kit have a stable and superior performance at higher temperatures.
二、產品說明
Template Total RNA, synthetic RNA transcript or poly(A)+mRNA, or the RNA should be avoided for cross-contamination with DNA.
Primer Selection Primer amounts recommended for efficient cDNA synthesis are 2.5 uM of oligo(dT) (anneal to the 3’-poly(A) + mRNA) or 2.5 ng/ul of random primers (anneal at non-specific sites of RNA templates), or 2 uM of genespecific primers per 20 ul reaction.
三、RScript cDNA 合成操作方法 (若有不同依原廠網站公告為主)
Reaction Setup cDNA Synthesis
1. For each 20 ul cDNA synthesis reaction, assemble the following in a PCR tube. Keep it on ice just prior to use.
*RNA template, primer and 2X Sharp Reaction Mix need to be premixed and heated at 65o C for 5 minutes in advance. After RNA has been pre-heated and incubated on ice bath at least 1 minute, add other components according to the table.
2. Mix the reaction solution gently by pipetting.
3. Cap the tubes and place them in the temperature-controlled water bath or heat blocks. Incubate the tubes at 55o C for 50 mins for the extension step. The 42°C – 60°C temperature range may be the optimal temperature for extension.
4. The reaction tube from the Step 3 must be incubated at 70o C for 15 minutes for inactivating the Reverse Transcriptase before amplification.
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