Ligation of the oligonucleotide will generate a HindIII site at the 3′-end of cDNA because of the presence of the poly(A) tail (Dorssers & Postmes, 1987). Inactivate the ligase by heating at 65°C for 15 min. cDNA is then digested with the restriction enzymes, HindIII and EcoRI, precipitated with ethanol and dissolved in 20 µl TE (pH 8.0).

Thus HindII and HindIII are two enzymes from Haemophilus influenzae strain Rd. Restriction endonuclease and modification methyltransferases (ENases and MTases) are sometimes distinguished by the prefixes R.EcoRI and M.EcoRI, but the prefix is commonly omitted if the context is unambiguous. The current convention for ENase naming omits ...

2017年7月1日 · For some endonucleases, including HindIII and DpnII, ligation of the 2 blunt ends generates a new digestion site that can be used to quantify the ligation and fill-in efficiency (NheI for HindIII and ClaI in the case of DpnII; Fig. 1 C, Fig. 2 B). After PCR amplification of a specific ligation product, the PCR product is digested with the ...

2012年11月1日 · The HindIII enzyme recognizes the sequence: 5′-AAGCTT-3′ and cleaves the DNA, leaving a 5′ overhang of: 5′-AGCT-3′. This cleavage provides a template for labeling the restriction fragments with biotin-14-dCTP, which will allow for the enrichment of Hi–C ligation products formed from crosslinked restriction fragments.

The mtDNA from maize with normal or fertile (N), T, C, and S cms cytoplasms have been studied by restriction enzyme fragment analysis (Levings and Pring, 1976, 1977; Pring and Levings, 1978). Four different restriction enzymes, Hind III, Eco RI, Bam I, and Sal I, were used in these analyses. Each restriction enzyme has a different specific ...

1977年9月1日 · The restriction sites on the λ genome between the left vegetative end and the b2 region were mapped entirely by successive digestion experiments. The restriction fragment map for the right arm of λ may be found in the accompanying paper (Robinson and Landy, 1977).

1983年10月1日 · Restriction enzyme X cleaves the DNA at one site, generating two fragments of size 200 and 800 base pairs. As depicted in Figure 1 A, the fragmentation pattern of X is used arbitrarily to assign the relative left-right orientation. Digestion of the DNA by restriction enzyme Y also results in two fragments of size 350 and 650 base pairs.

1977年9月1日 · The site-specific restriction endonucleases isolated from Hemophilus influenzae strains Rc (HincII) and Rd (HindII+III), and Hemophilus parainfluenzae (HpaI) were used to digest bacteriophage λ DNA into 34, 40, and 15 specific fragments, respectively. The sites cleaved by each of these enzymes were localized on the λ physical map and the ...

1996年12月1日 · The number of restriction enzyme sites varied from 3 for NotI to 10 for HindIII and KpnI, and the sizes of restriction enzyme fragments varied from 0.4 to 26 kb. The genome size of the AAV type 8 as calculated from the RE cleavage ranges from 44.4 kb (SpeI) to 45.0 kb (KpnI) with an average of 44.7 kb for all eight REs.

(A) To determine the location and number of restriction enzyme sites, a segment of DNA is digested with a restriction enzyme. In this example, a piece of DNA 5000 bp in length was cut with BamHI to give three fragments: 3000 bp, 1500 bp, and 500 bp. The figure shows the three possible arrangements of these three fragments.