recombinant dna part d

... and thus overcome the drawbacks of the filamentous phage vectors described above. 2°,2°a 2t D. Denhardt, D. Dressier, and D. S. Ray (eds.), "The Single-Stranded DNA Phages." Cold ... strands packaged by IKe and Ff, different primers must be used for sequence analysis, hybridization studies, or site-directed mutagenesis of the DNA inserts. The ssDNA packaged under the di...

Ngày tải lên: 11/04/2014, 10:26

... concentration of DNA ligase is needed for complete joining than if DNA were not incu- bated with phosphatase. Cohesive ends can be added to blunt-ended DNA molecules by liga- tion with synthetic DNA linkers. ... to PstI-cleaved pBR322 DNA and dC tails to the DNA to be inserted, anneal, and transform E. coli with the DNA. PstI sites are reconstituted on both sides of t...

Ngày tải lên: 11/04/2014, 10:25

... purity of the DNA substrate. Improp- erly prepared DNA samples will be cleaved poorly, if at all, producing partially digested DNA. In addition to DNA purity, other DNA- associ- ated parameters ... required to digest completely 1 /~g of lambda DNA under standard reaction conditions. ' All enzymes and DNAs were from Bethesda Research Laborato- ries, Inc. d Linear form II...

Ngày tải lên: 11/04/2014, 10:25

... nematode DNA added as long as the DNA concentration is low. The ligation reaction is saturated with a greater than 2.0-fold molar excess of insert DNA to vec- tor DNA (0.5/~g insert DNA per ... and is considered to be a "pilot" protein. The viral single-stranded DNA (ss -DNA) is converted into a double-stranded circular form (RF DNA) without the synthesis of any v...

Ngày tải lên: 11/04/2014, 10:26

... of RNA Strand The HindlII-cut, C-tailed cDNA : mRNA-vector is cyclized by DNA ligase and the oligo(dG)-tailed linker DNA that bridges the C-tail and the HindlII end of the hybrid-vector. The ... preparations by endonuclease specific for double- or single-stranded DNA can be detected by digestion of supercoiled pBR322 or single-stranded ~bX174 DNAs under the conditions specified b...

Ngày tải lên: 11/04/2014, 10:26

... 10.0 mM ddCTP 8 10.0 mM ddGTP" 16 10.0 mM ddATP ~ 1 TE buffer 1000 1000 1000 500 The exact ratio of dNTP to ddNTP may need to be determined empiri- cally and adjusted accordingly; i.e., ... pairs) of the eight DNA fragments produced by NotI cleavage of Adeno-2 DNA and the sizes of the four DNA fragments produced by SfiI cleavage of Adeno-2 DNA are listed in Table I. Rel...

Ngày tải lên: 11/04/2014, 10:26

... flour embedded in agarose probed with pAW173. Agarose-embedded rye flour was lysed for 48 hr and the released DNA was digested with various restriction enzymes. Lane 1, unrestricted DNA; lane ... double band coincides with the accessibility of the DNA preparation to specific endonuclease digestion. The hybridization with the radiolabeled probe pAW173 of the DNA digested with Hind...

Ngày tải lên: 11/04/2014, 10:26

... Plasmid DNA from Bacillus brevis Cells The method of Birnboim and Doly 12 can be used to obtain plasmid DNA. This method can be scaled up to obtain large amounts of plasmid DNA and DNA can ... NaCI, and I0 mM EDTA), 5/zl of 10% (w/v) sodium dodecyl sulfate (SDS), and 40 ~1 of distilled water. Incubate 15 min at 70 °, extract with phenol and chloroform, and precipitate the DNA a...

Ngày tải lên: 11/04/2014, 10:26

... better sequencing ladders. Protocols for se- quencing double-stranded amplified DNA (dsDNA) by the dideoxy method usually involve a DNA denaturation step followed by a rapid annealing to a specific ... Single-stranded DNA sometimes migrates ahead of the dsDNA, but it does not stain well with ethidium bromide. To determine the effect of limiting the amount of one primer on the yie...

Ngày tải lên: 11/04/2014, 10:27

... the screened cDNA library, DCoH ⁄ PCD and the DCoH ⁄ PCD transcript are hardly detected [12,15]. PDIP46 was first identified in the yeast two-hybrid assay designed to find molecular partners of ... yeast two-hybrid system, we identified the PDIP46 ⁄ SKAR protein encoded by the polymerase (DNA- directed), delta interacting protein 3 (POLDIP3) gene as a bind- ing partner of the human ER. PDIP46⁄ SK...

Ngày tải lên: 19/02/2014, 05:20