recombinant dna part f

... BSA (nuclease free) 5 raM dithiothreitol (0-100 mM NaCI) 2. Replace buffer with two volumes of fresh buffer. Methylation is performed in a volume twice that of the beads in buffer containing ... of ~bX174 RF I DNA in the standard restriction endonuclease assay buffer in a volume of 0.05 ml. The supercoiled ~bX174 DNA has been substituted for Adeno-2 DNA because neither NotI nor...

Ngày tải lên: 11/04/2014, 10:26

... of genes, the sequence analysis of DNA, and the identi- fication of regulatory elements for DNA replication and transcription depend on the availability of pure specific DNA segments. The DNA ... for a given DNA sequence by nucleic acid hybridization. 43 Second, in vitro packaging of recombinant DNA molecules provides a very efficient means of infecting bacteria with them....

Ngày tải lên: 11/04/2014, 10:25

... incubation of an appropriate DNA with various amounts of the respective endonuclease in the standard reaction buffer, in core buffer, or in core buffer supplemented with NaC1. The standard buffer ... other hand, effective. Citrate and other biological buffers that chelate Mg 2+ cannot be used. The selected buffer concentration must be sufficient to maintain the proper pH of the final r...

Ngày tải lên: 11/04/2014, 10:25

... LAMBDA VECTORS WITH SELECTION FOR INSERTS 13 FIG. 4. Fractionation of partially digested nematode DNA. Nematode DNA (N2 DNA) was prepared from frozen animals purified by flotation on sucrose? The ... synthesis of any viral product. The synthesis of the first RF DNA (parental RF) is an interesting step, because by this means the virus passes from a passive into an active life form....

Ngày tải lên: 11/04/2014, 10:26

... of ssDNA of (recombinant) plas- mids. Cloning of the viral strand (+) origin plus morphogenetic signal of either Ff or IKe into a plasmid diverts the plasmids upon superinfection to the Ff ... vitro recombination of the largest BanI fragment of RF of the Ff phage R4087°a and the smallest BanI fragment of Mike RE. 69 The major advantage of the introduction of this deletion in ......

Ngày tải lên: 11/04/2014, 10:26

... /zl of 10x terminal transferase buffer, 1 /zl of 1 mM DTT, 20/xg of KpnI-digested DNA (19 pmol of DNA ends), 1/xl of 2.5 mM [3H]dTTP (250-500 dpm/pmol), and 20 units of terminal trans- ferase. ... 1 2 FIG. 3. Effect of different wash conditions. Five (lanes 1) and 10/zg (lanes 2) amounts of BamHI-digested C57BL/6 liver DNA were prepared and hybridized as described in Fig. 1. Aft...

Ngày tải lên: 11/04/2014, 10:26

... pellet, forcefully pipette TEX or buffer into the tube (the force of the buffer injected into the tube is sufficient to resuspend the microbeads completely), centrifuge for 1 min in a microfuge ... pellet from the buffer, hold the Eppendorf tube up to a light to make the pellet/buffer boundary visible by the difference in diffraction. The buffer remaining in the microbead pellet after .....

Ngày tải lên: 11/04/2014, 10:26

... insertion of foreign ORF DNA fragments in plFF8 has a more pronounced effect on the Lac ÷ phenotype. Consistent with this notion, insertion of a 1.6-kb pal cDNA fragment in plFF8 produced ... of sequencing data. As with other ORF vectors, the proper function of plFF8 requires that the lacZ-encoded part of the fusion protein retain enzymatic activity. In plFF8, the foreign DNA...

Ngày tải lên: 11/04/2014, 10:26

... bromide. To determine the effect of limiting the amount of one primer on the yield of dsDNA and ssDNA, we conducted the experiment shown in Fig. 3. Different amounts of one primer (50, 20, 15, ... (1991). FIG. 4. Sample sequencing ladders obtained from PCR-amplified ssDNA. (A-C) Amplifi- cations from Mus spretus genomic DNA; (D -F) amplifications from Drosophila melanogaster genomi...

Ngày tải lên: 11/04/2014, 10:27

... large amounts of a particular sequence • The number of molecules of the DNA fragment between the primers increases about 2-fold with each cycle. • For n = number of cycles, the amplification is ... DNA fragment) than YACs • Extensively used in genome projects BAC vectors for large DNA inserts Cut with restriction enzyme E, remove “stuffer” Ligate to very large fragments of genomic...

Ngày tải lên: 13/03/2014, 18:39