Restriction Landmark Genomic Scanning (RLGS) by Yoshihide Hayashizaki (auth.), Yoshihide Hayashizaki M. D., PDF
By Yoshihide Hayashizaki (auth.), Yoshihide Hayashizaki M. D., Ph. D., Sachihiko Watanabe Ph. D. (eds.)
Restriction Landmark Genomic Scanning (RLGS) is a brand new multiplex approach for simultaneous research of greater than 3,000 genome loci. Written by means of the inventor of RLGS, Yoshihide Hayashizaki, and his co-workers, this is often the 1st guide in this approach. RLGS is a strong strategy with huge, immense capability for organic (animal and plant sciences), and biomedical study. Yielding effects a lot quicker than traditional recommendations, RLGS is very suited for the identity of the chromosomal situation of the genes implicated in inherited ailments, and for the genetic disturbances found in cancerous tissue.
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Extra info for Restriction Landmark Genomic Scanning (RLGS)
3 Practical Application of the PCR-Mediated Method . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 Breakthrough in the Development of RLGS Spot Cloning In order to realize the potential of this technique, it is essential to have a direct and simple method for recovering individual loci of interest. A direct method for recovering landmark spots or sites is also essential for transferring the information to other laboratories and for identifying the gene functions that may be associated with these sites.
Hayashizaki - Notl adaptor (consisting of 5'-ACGCCAGGGTTTTCCCAGTCACGACGC-3' and 5'-pGGCCGCGTCGTGACTGGGAAAACCCTGGCGT -3') - Hinfi adaptor (consisting of 5'-pANTCTGTACTGCACCAGCAAATCC-3' and 5' -GGATTTGCTGGTGCAGTACAG-3') - P77 (PCR primer), 5'-AGGGTTTTCCCAGTCACGACGCGG-3' - Ad2-2 (PCR primer), 5'-TTGCTGGTGCAGTACAGANTC-3' - P8 primer (Toyobo) Stock - T AE buffer buffers and solutions - TE buffer - T 10E0. 4), lOOmM MgC12 , lOmM DTT] - lOx ligation buffer (for E. 5mM) - lOx kination buffer [SOOmM Tris-HCl, IOOmM MgC12 , SOmM DTT] - lOOmM ATP - 50% polyethylene glycol6000 (PEG) - 3M sodium acetate - 1% linearly polymerized acrylamide 4 RLGS Spot Cloning 51 Recovering DNA from the Punched-Out Target Spot Recovering of DNA 1.
60. Extract with PCI. 61. Ll (1% linearly polymerized acrylamide is used as carrier). Incubate at -80°C for lSmin. 63. Centrifuge at 15 000 rpm for 30 min. 64. Ll TE. Preparation of Noti-Pstl Vector 65. Digest the pBS II with Notl and Pstl. 66. Electrophoresis; recover the vector fragment (dephospholylation was not carried out). ll) 50% PEG 6000 Total 68. Ligate overnight at 18 °C. 69. Transform. ll 48 S. Watanabe andY. Hayashizaki RLGS gel Hurfl Nod p Hurfl Nod ligation with adaptors Hurfl Not.!
Restriction Landmark Genomic Scanning (RLGS) by Yoshihide Hayashizaki (auth.), Yoshihide Hayashizaki M. D., Ph. D., Sachihiko Watanabe Ph. D. (eds.)