The DNA pellet was subsequently re-suspended in 4 ml of 1× TE (pH 7.4) and proteinase K digested by the use of 400 µl of 10X buffer (240 mM EDTA, 750 mM NaCl, pH 8.0), 400 µl of 10% SDS and 20 µl of proteinase K (20 mg/ml), and samples were incubated overnight at 48°C with constant shaking.
Help readers to remember and digest concepts with the use of numbers or bullets.
Digest reactions included the use of trypsin digestion alone; trypsin digestion followed by Glu-C digestion; Asp-N digestion followed by Arg-C digestion; or Asp-N/Arg-C digestion followed by Glu-C digestion.
The basic idea of shotgun proteomics is to digest the proteome of interest using specific enzymes and identify the resulting peptides by sensitive and accurate MS analyses.
The fourth type uses proteases and hydrolytic enzymes; these enzymes are used to digest the proteins of destroyed bacteria.
Successful FISH analysis depends on a pretreatment protocol that is consistent in the use of digesting agents (sodium thiocyanate, pepsin or proteinase K) as well as appropriate conditions (concentration, temperature and time of incubation) to unmask the target DNA and optimize hybridization.
Heparitinase I (EC4.2.2.8), and chondroitinase ABC (EC4.2.2.4) (both from Seikagaku, Tokyo, Japan) were used to digest the side chains of the heparan sulfate and chondroitin sulfate proteoglycans, respectively.
Trypsin from bovine pancreas (Sigma-Aldrich) was used to digest the C2-block of C2SH8C2, C2SH16C2, and C2SH24C2.
The following restriction endonucleases were used to digest the genomic DNA of PaP1 or PaP3 in 20 μL reaction systems according to the manufacturer's instructions: PauI; VspI; AatII; SpeI; and EcoRI (New England Biolabs, Ipswich, MA, USA).
A MAC combines the use of a message digest with a secret key known only to two users.
