recombinant dna part i

... ssDNA is visible after the bromphenol blue has migrated about 2 cm as a discrete fraction migrating ahead of the dsDNA. If a ssDNA fraction is visible by ethidium staining, the asymmetric ... asymmetric, or unequal, ratio of the two amplifica- tion primers is used in the PCR 6 (Fig. 1). During the first 20-25 cycles dsDNA is generated, but when the limiting primer is exhausted ssDNA is ....

Ngày tải lên: 11/04/2014, 10:27

... efficiency as supercoiled plasmid DNA, it can give rise to clones containing vehicle plasmid rather than a recombinant. Inactivation of a genetic marker is not an infallible indication of DNA ... chloramphenicol resistance, by DNA insertion at their single EcoRI sites (Table I) . Insertional marker inactivation is a useful feature in a cloning vehicle. However, it is not essential...

Ngày tải lên: 11/04/2014, 10:25

... RECOMBINANT DNA [1] r~ o [-, z o z o o Z 0 Z e ,q I I I I I I ~ I I.~'~ I I- I& quot;~.~ t ~ I I I ,~ I~ o,~ I~ =~ ~- I I I I,~,~ I 0 I~ I~ I I I I I I I~ ° I I I~ I oA ... oA O0 [1] USE OF TYPE II RESTRICTION ENDONUCLEASES 7 I ~ I I I ~~.~.~1 I I I I ~-~-~ I I I °~ I I I I~ I I ~ I ~ ~ v. f g. ~D ~ he3...

Ngày tải lên: 11/04/2014, 10:25

... (XbaI) [int-clII]cI857 hhsbaml°b189(int(linker)ninL44 D EcoRI, BarnHI, 9-22 A[sHindlII3-sHindlII5],EtrpE)int(linker) SalI A[in t-c III]KH54a RI4°nin 5s RI5 ° control, and a 9-11 kb right ... RI4°nin 5 XhoI linker in 1059 BamHI sites D BamHI, XhoI 9-22 XbaI linker in 2004 BamHI sites C XbaI 7-20 hhsbaml°b189att int (XbaI)ninL44 C XbaI 5-18 A[sHindlII3-sHindlII5]~bio 256...

Ngày tải lên: 11/04/2014, 10:26

... RNA primer for initiation I IJTJ of - strand sunthesis I J J I& quot; niok site for gone II protein I II d / (initiation of + strand synthesis) tv | - ^ I . morphogenesis signal + ... virion proteins, V is an ssDNA binding protein, X is probably involved in repli- cation, and II creates a site-specific (+) strand nick within the IG region of the double-stranded replicati...

Ngày tải lên: 11/04/2014, 10:26

... the SDS/EDTA solution and 6O 2 < 45 n- i I r I i I I .=20 lO ~c i I i I i I L I i I I i 0 J I I I I I I 0 10 20 30 ( 10 20 30 Time (min) Time (min) FIo. 4. Time course of poly(dT) ... cDNA containing multiple HindlII sites/ Using the reaction conditions described below, the HindlII cleavage is often relatively poor (30-50% digestion). This may be d...

Ngày tải lên: 11/04/2014, 10:26

... characterize at least two different mismatch ligations: GGCACC ~ GGCACC III1 IIII IIII IIIII CCACGG 1' CCATGG 1' It is worthwhile to note that these ligation products did not contain ... BanI fragments required for replication and ampicillin resis- tance. In a second experiment using another multilinker variant of pHK402, pHK400, containing an additional HgiCI (KpnI) site w...

Ngày tải lên: 11/04/2014, 10:26

... replication (like that of the small animal DNA viruses simian virus 40, polyomavirus, or bovine papillomavirus type 1), which is included if replication is desired in animal cells providing the viral ... molecular biology. The simplest vector constructions contain a plasmid origin of replication, one or two antibiotic resistance genes, which enable selection in antibiotic-containing me...

Ngày tải lên: 11/04/2014, 10:26

... I I I ! I I i I i I I i ! I ! I I I ! I ! I SP6 promoter A GCTI'GA GTATTCTITI'A GTGTG A G CTA A ATAGCTFG 6C GTAATC ATG GTC AT '~ Plac I I I I I | I I I | I I I I I | I I ... used and in-frame cloning is facilitated by size randomizing the DNA insert prior to cloning. We discuss here a novel in-frame cloning principle that simpli...

Ngày tải lên: 11/04/2014, 10:26

... join DNA: homopolymer tails Alternate method to join DNA: linkers Introduction of recombinant DNA into living cells via vectors • Autonomously replicating DNA molecules – (have an origin ... procedures to isolate a gene that represents as little as 1 part in a million of the genetic material in an organism. • DNA from the organism of interest is divided into small p...

Ngày tải lên: 13/03/2014, 18:39