Can In-Fusion be used to clone a microRNA (miRNA) precursor

-Sequences for microRNA precursors and flanking genomic DNA can be obtained from a number of public databases, including GenBank and EMBL-Bank. The UCSC Genomic Bioniformatics Site hosts an easy-to-navigate genomic database which tracks miRNAs. The Sanger Institute hosts miRBase, a compilation of known miRNA sequences. -100–300 bp of DNA flanking the miRNA precursor is amplified from genomic DNA [...]

Can In-Fusion be used to clone shRNA (small hairpin RNA)?

An shRNA double-stranded DNA oligonucleotide (?50 bp) can be cloned via In-Fusion technology into a linearized shRNA expression vector. -For In-Fusion cloning of short synthetic oligos (between 50 and 150 bp), the suggested oligo-to-vector molar ratio is 5–15:1. Depending on the oligo length, the optimal ratio must be determined empirically. -High-quality, non-phosphorylated oligonucleotides [...]

What's In-Fusion's compatibility with site-directed mutagenesis?

In-Fusion Cloning allows single or multiple base changes, deletions, and insertions through the use of inverse PCR. Please note this application relies on high-fidelity PCR. It is essential to use a PCR polymerase with high proofreading activity, such as PrimeSTAR Max DNA Polymerase. This polymerase is highly robust and accurate, enabling amplification of up to 6 kb of human genomic DNA, 10 kb [...]

What's In-Fusion's compatibility with multiple-fragment cloning?

We have successfully tested multiple-fragment cloning with up to five inserts. (See Figure and Table below for cloning schematic and colony screen results, respectively.) Primer design for multiple-fragment cloning can be done with our online Primer Design Tool. (The Primer Design Tool is compatible with Mozilla Firefox or Google Chrome web browsers, but not with Internet Explorer.) Please note [...]

What's In-Fusion's compatibility with large inserts?

This technology has been optimized for cloning large fragments. DNA inserts up to 15 kb have been successfully cloned into pUC19 using In-Fusion Cloning (results confirmed by colony PCR screening).In-Fusion Cloning [...]