Cohesive 5 overhang 3 overhang cloning Polylinker or

= מתלכד Cohesive 5’ overhang 3’ overhang

cloning

Polylinker or Multiple Cloning Site (MCS)

Plasmid cloning •

Taq polymerase: is a thermostable DNA polymerase named after thermophilic bacterium Thermus aquaticus from which it was originally isolated by Thomas D. Brock in 1965. Taq polymerase

PCR – polymerase chain reaction 30 sec for 1 Kb = elongation

PCR first PCR-2 מומלץ לצפייה In vitro mutagenesis / site-directed mutagenesis – Creating transgene mouse

The PCR product on the gel

Site-directed mutagenesis

Noam Shomron / Tel Aviv University / The RNA World / January 2010 Deep-sequencing 454/Roche, Solexa/Illumina, ABI/Solid

Noam Shomron / Tel Aviv University / New Horizons in RNA processing / Nov-Dec 2009 OLD NEW $3 billion 10 years 20, 000 BACs 100 kb > 2 -3 kb vectors

Principle of Microarray (Chip) Assay Prehybridization Posthybridization Synthetic DNA probes Probes with hybridized DNA

microarray Genome microarray

Ch. IP-on-chip Antibody Binding sites TF TF Genomic Arrays

Ch. IP-seq in a nutshell

Native gel

: 1 תמונה רצף גנומי חלקי של הגן survival of motor neuron 1. (SMN 1) Exon – red upper case Intron – black lower case Amino acids –under the center of the codon

אתרי חיתוך של שלושה אינזימי רסטריקציה : 3 תמונה Products generated by restriction enzymes Cohesive ends Eco. RI Bam. HI 5’ GAATTC 3’ 3’ CTTAAG 5’ 5’ GGATCC 3’ 3’ CCTAGG 5’ Xho. I 5’ CTCGAG 3’ 3’ GAGCTC 5” 5’ G AATTC 3’ 3’ CTTAA G 5’ 5’ G GATCC 3’ 3’ CCTAG G 5’ 5’ CTCGA 3’ G G 3’ AGCTC 5’