Question: Why does gel purified DNA template often yield poor results in transcription reactions?

Answers: Low quality DNA templates are often associated with poor transcription reactions because the gel purified DNA are cross-linked by ethidium bromide and contaminated with traces of impurities from the agarose. DNA templates from ethanol precipitation can not be used in the transcription reaction because they also precipitate free nucleotides, proteins and salts in the reaction solution. 

NeuBiogene's Transcript Purification Kit (Cat# TP-25) provides a fast and efficient way to completely separate synthesized RNAs from free nucleotides, proteins and salts.  Purified RNA transcripts can be immediately used for functional studies and hybridization experiments.

Question: Why does the capping transcription kit only give you about 2-3 µg of the total capped mRNAs in a 20 µl reaction? Do you know that you can increase your results by nearly 5 times in the same volume of reaction?

Answers: In our capping kits (Cat# RMP-25), a special solution was added in the transcription system so that the total yield of capped mRNA can reach >15 µg in a 20 µl of transcription reaction.  Also, the capping efficiency is >90%.

Question: What is the difference between miRNA and siRNA?

Answers: miRNA is a single stranded RNA with a hair-pin structure and siRNA is a double-stranded RNA without hair-pin structure. One major difference between the two is that miRNA is loaded into Ago1 and siRNA is loaded into Ago2. Ago1 allows mismatches, while Ago2 allows only perfect short RNA - mRNA pairs.

Question: Do you know that you can isolate miRNA just like you isolate DNA or RNA?

Answers: Neubiogene’s MiniExpress Plasmid Isolation (RNase Free) kit lets you isolate and enrich miRNAs or siRNAs from tissue samples or cultured cells transfected with siRNA or miRNA-expressing vectors. No acrylaminde gel or SDS PAGE is necessary. The recovery rate of enriched siRNAs or miRNAs is much higher than PAGE gel based isolation and the total time for enriching small RNAs is also much shorter than PAGE gel based method.  

Question: Do you know that you can use RNase A to remove the high background and non-specific binding of propidium iodide?

Answers: Before you add Propidium Iodide dye to the cells with the culture medium, you can add RNase A (1 ul per ml culture medium, Cat# RNase A-50) and incubate for 15 min at 37C. RNase A treatments will remove the RNAs released from the dead and dying cells which can bind to propidium iodide and give you the false positive and high background.