Lambda phage dna
Lambda phage DNA is a linear double-stranded DNA molecule that serves as the genetic material of the lambda bacteriophage. It has a size of approximately 48.5 kilobase pairs and contains the necessary genetic information for the phage's replication and infection of host bacteria.
Lab products found in correlation
8 protocols using lambda phage dna
Biotin-Labeled dsDNA Amplification from Lambda Phage
Histone Exchange Reaction Assay
Histone Exchange Assay for SWR Chromatin Remodeling
DNA-Protamine Binding Characterization
DNA of varying lengths [L = 25 nm (75 bp), 50 nm (150 bp), 105 nm (309 bp) and 217 nm (639 bp)] was made via Polymerase Chain Reaction (PCR) using standard protocols (29 (link),32–34 (link)), lambda phage DNA (New England Biolabs, N3011L) as a template, and LA Taq DNA polymerase (TaKaRa Bio, RR002). To biochemically tether the DNA to the surface of the cover slip and the streptavidin-coated polystyrene bead, one primer was labelled with a digoxigenin molecule, while the other was labelled with a biotin molecule (Integrated DNA Technologies). To purify the DNA, we performed gel electrophoresis using orange loading dye (B7022S, New England Biolabs) and standard protocols for short DNA molecules (35 ), then purified the DNA via gel extraction (QIAquick Gel Extraction Kit, Qiagen). We quantified the final concentration using a spectrophotometer (Nanodrop One, ThermoFisher Scientific).
DNA Labeling and Nanosphere Characterization
Suncoast yellow fluorescent polymer nanospheres (excitation/emission maxima 540/600 nm) of nominal size (0.19 μm; 2.653 × 1012 nanospheres ml−1) were purchased from Bangs Laboratories, Inc. (Fishers, IN, USA). The nanospheres were diluted with 70% glycerol in 10 mM TRIS buffer (pH 8) to yield a concentration of 1.5 × 106 nanospheres ml−1.
Biotinylated DNA Synthesis via PCR
∼5 kb DNA
with biotin attached at one end, we performed PCR according to the
manufacturer’s recommendations (PCR Extender System, QuantaBio,
Beverly, Massachusetts) using Lambda phage DNA as a template (New
England Biolabs Inc., Ipswich, Massachusetts), a biotinylated primer
and an unmodified primer (forward 5′-biotinTEG-CTGATGAGTTCGTGTCCGTACAACTGGCGTAATC-3′,
reverse 5′-GTTTGTACTCCAGCGTCTCATCTTTATGCGCC-3′,
Integrated DNA Technologies). This produced a single band in a conventional
agarose gel electrophoresis experiment at the expected 5031 bp. The
cleaned PCR product was combined with biotin-plugged divalent streptavidin,
where the oligonucleotides of the two plugging molecules had been
cleaved off with 5 min UV irradiation, and incubated overnight in
100 mM sodium phosphate buffer pH 6.5. Products of the incubation
were visualized on a 0.5% agarose gel using staining with ethidium
bromide.
Characterization of SYBR Gold Dye
Fabrication of Semiflexible Magnetic Filaments
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!