For protein tagging and labeling of intracellular structures study, miRFP670nano was amplified, digested with restriction enzymes, and then swapped with miRFP703 either as C- (for α-tubulin and clathrin) or N-terminal fusions (for keratin, α-actinin, LifeAct, EB3, myosin, vimentin, clathrin, LAMP1, and H2B 3.
Each cDNA was amplified, digested, run on gel, and quantified three times (n = 9).
Genomic DNA was prepared from cell pellets, PCR amplified, digested, and hybridized onto GMAP arrays (Affymetrix Inc).
The AtCTF7 promoter (1.3 kb upstream of the ATG) was amplified, digested and cloned into the 35S AtCTF7∆B construct.
cDNA generated by reverse transcription was amplified, digested with Sfi 1A and Sfi 1B, and size fractionated.
Subsequently, the cDNA was amplified, digested with one of two restriction enzymes, and the appropriate Illumina-based sequencing adapter appended to the digested molecules by ligation.
Mkrn1 protein-coding cDNA was amplified, digested with PvuII to generate blunt ends, and ligated into the pCAGGS::FLAG vector, immediately downstream from the FLAG epitope to yield the pCAGGS::N-FLAG MKRN1 N-FLAG MKRN1
The 5′ and 3′ halves of CD81 were amplified, digested and ligated into pJET2.1 to produce full-length CD81 ORFs encoding the desired mutations that were transferred into pTRIP-AcGFP or pTRIP-DsRED plasmids and sequenced.
For DR5::GUS vector construction, the DR5 element (Ulmasov et al. [1997]) coupled to the CaMV 35S minimal promoter was amplified and digested at the site of Sal I and Bam H I, and inserted into pBI101.3, which carries the structural gene for GUS and the terminator sequence of the nopaline synthase (NOS) gene.
A region in the 16S rRNA was amplified and digested with the endonucleases DdeI, BsrI, and TaqI.
To determine if this polymorphism is present in our fly population, a 201 bp fragment from H1 coding sequence encompassing the position 688 was amplified and digested with SfcI.
