DNA Technology

• COPPIED/CLONED
• MADE INTO LIBRARIES
• SEQUENCED

CLINING PERMITS PRODUCTION OF MULTIPLE COPIES OF A SPECIFIC GE OR DNA SEGMENT

• POLYMERASE CHAIN REACTION(PCR) COPIES SPECIFIC DNA SEQUENCE
• REACTIONS THAT CAN AMPLIFY TARGET DNA SEQUENCES PRESENT IN VERY SMALL QUANITIES

TWO OLIGONUCLEOTIDE PRIMERS

ONE COMPLEMENTARY TO THE 3' END OF ONE STRAND THE DNA TO BE AMPLIFIED

ONE COMPLEMENTARY TO THE 3' END OF THE OTHER STRAND

PRIMERS ANNEEAL TO DENATURE DNA/ COMP STRANDS ARE SYNTHESIZED BY HEAT STABLE DNA POLYMERASE

DENATURATION

ANNEALING

EXTENSION

methods for preparing well defined/gene size pieces of dna in multiple identical copies

CUT NA MOLECURES AT A LIMITED NUMBER OF SPECIFIC DNA SEQUENCES CALLED RESTRICTION SITES

USUALLY MAKE MANY CUTES IN DNA MOLECULE-YIELDING A SET OF RESTRICTION FRAGMENTS

STAGGERED WAY- PRODUCING FRAGMENTS WITH STICKY ENDS THAT CAN BOND WITH COMPLEMENTARY STICKY ENDS OF OTHER FRAGMENTS

ENZYME THAT SEALS THE BONDS BETWEEN RESTRICTION FRAGMENTS

ENZIMES CUT AT RESTRICTION SITE

DNA FRAGMNT FROM ANOTHER ORGANISM IS ADED BASE PAIRING ON STICKY ENDS PRODUCE VARIOUS COMBINATION

DNA LIGASE SEALS THE STRANDS

SEPERATES DNA RESTRICTION FRAGMENTS OF DIFFERENT LENGHTS

LONGER(SLOWER FRAGMENTS) TOP

SHORTER(FASTER FRAGMENTS) BOTTOM

USEFUL FOR COMPARING TWO DIFFERENT DNA MOLECULES SUCH AS 2 ALLELES FOR A GENE

PCR CAN CLONE SHORTSEGMENTS OF DNA BUT NOT ENTIRE GENOMES

GENOMIC LIBRARY CONTAIS AT LEAST ONE COPY OF ALL SEQUENCES IN THE GENOME

LIBRARIES CAN BE MADE BY CUTTING GENOMIC DNA WITH RESTRICTION ENZYMES AD LIGATING FRAG INTO VECTOR

INVOLVES MARKING THEN SORTING CHROMOSOMES

SUBGENOMIC FRACTIONS SUCH AS SINGLE CHROMOSOMES CAN BE SORTED USING THIS TECHNIQUE

CONTAINS COMPLEMENTARY DNA COPIES MADE FROM MRNAS

REPRESENTS GENES THAT ARE TRASNCRIPTIONALLY ACTIVE AT THE TIME THE CELLS WERE COLLECTED FROM MRNA ISOLATION

ISOLATING MRNA FROM CELLS

• SYNTHESIZING COMP. DNA USING
• REVERSE TRANSCRIPTASE

CLONING CDNA MOLECULES INTO VECTOR

• 2 MAIN TECHNIQS
• SCREENING

MAPPING

PROBES COMP. TO PART OF A GENE ARE USED TO SCREEN A LIBR. TO RECOVER CLONES OF A SPECIFIC GENE

CLONES GROWN ON AGAR PLATES TO PRODUCE COLONIES

COLONIES SCREENED BY TRASFERRING BACTERIAL COLOLINES FROM PLATE TO FILTER AND HYBRIDIZIN FILTER WITH A NUCLEIC ACID PROBE TO THE DNA SEQUENCE OF INTEREST

COLONY CORRESPONDING TO THE ONE ON THE PROBE IDENTIFIED ON THE FILTER IS IDENTIFIED AND RECOVERED

A RESTRICTION MAP ESTABLISHES THE NUMBER AND ORDER OF RESTRCITION SITES AND THE DISTANCE BETWEEN RESTRICTION SITES ON A CLONED DNA SEGMENT

MOST COMMON METHOD OF DNA SEQUENCING IS DIDEOXY CHAIN TERMINATIO SEQUENCING DEVELOPED BY SANGER

DNA SEQUENCES IS DENATURE INTO SINGLE STRANDS

SINGLE STRANDS ARE MIXED WITH PRIMERS THAT ATTACH TO THE 3' END OF DNA

• SAMPLES GO INTO 4 TUBES W/ DATP DGTP DCTP OR DTTP
• TUBES LABELED DDEOXYNUCLEOTIDE (HAVE 3 OH)
• DNA SYNTHESIS TAKES PLACE AND PRIMES ARE EXTENDED

ALTERATION OF AN AFFICTED INDIVIDUALS GENES

"NORMAL" NON DISEASE GEN IS INTRODUCED INTO THE AFFECTED INDIVIDUALS GENOMES USING A VECTOR

HOLDS GREAT POTENTIAL FOR TREATING DISORDERS TRACEABLE TO A SINGLE DEFECTIVE GENE

SPECIFIC PATER OF BANDS OF RFLP MARKERS ON A GEL

TRANSGENIC ANIMALS CONTAIN GENES FROM OTHER ORGANISMS

IS THE MOST COMMONLY USED VECTOR FOR INTRUDUCING NEW GENES INTO PLAN CELLS