miRcute miRNA Isolation Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Addition of Buffer MZ depends on monolayer area, not cell number as it will reflect on DNA contamination if not suffcient.	- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.

Upstream tips
- Addition of Buffer MZ depends on monolayer area, not cell number as it will reflect on DNA contamination if not suffcient.
Protocol tips
- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.

Publication protocol

miRNAs were isolated by miRcute miRNA isolation
kit (Tiangen, China). The extracted products were then
done reverse transcription (RT) reaction using miRcute
miRNA first-strand cDNA synthesis kit (Tiangen, China). The synthesized first-strand cDNA was kept at -20℃. All procedures were performed according to the instructions provided by the manufacturer. The miR-155 level was quantified by quantitative reverse transcription-PCR (qRTPCR) with 5s small nuclear RNA as an internal normalized reference. The qPCR was performed on ABI 7500 RealTime PCR System (ABI,USA) with miRcute miRNA qPCR detection kit (Tiangen, China), which included specific reverse primer for miRNAs. The qPCR reaction system contained 10ul 2×miRcute miRNA premix, 0.4ul miR-155 primer, 0.4ul reverse primer, 2ul ten fold serial dilution of miRNA first-strand cDNA and 7.2ul RNasefree water. The procedure for PCR was 94℃ 2 min; 94℃ 20s, 60℃ 30s, 72℃ 30 s, 45 cycles

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Cells - immortalized MDA-MB-157 using miRcute miRNA Isolation Kit from Tiangen.

Manufacturer protocol

Download the product protocol from Tiangen for miRcute miRNA Isolation Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Cells - immortalized MDA-MB-157 using miRcute miRNA Isolation Kit from Tiangen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.