DNA Technology

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Author:
datgrl_honey
ID:
115444
Filename:
DNA Technology
Updated:
2011-11-09 18:46:23
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DNA technology
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Dna Technology
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  1. DNA /TECHNOLOGY
    • COPPIED/CLONED
    • MADE INTO LIBRARIES
    • SEQUENCED
  2. DNA CLONING
    CLINING PERMITS PRODUCTION OF MULTIPLE COPIES OF A SPECIFIC GE OR DNA SEGMENT
  3. CLONING PROCESS
    • POLYMERASE CHAIN REACTION(PCR) COPIES SPECIFIC DNA SEQUENCE
    • REACTIONS THAT CAN AMPLIFY TARGET DNA SEQUENCES PRESENT IN VERY SMALL QUANITIES
  4. PCR REQUIRES
    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
  5. PCR
    PRIMERS ANNEEAL TO DENATURE DNA/ COMP STRANDS ARE SYNTHESIZED BY HEAT STABLE DNA POLYMERASE
  6. 3 STEPS PCR
    DENATURATION

    ANNEALING

    EXTENSION
  7. GENE CLONING
    methods for preparing well defined/gene size pieces of dna in multiple identical copies
  8. BACTERIAL RESTRICTION ENZYMES
    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
  9. MOST USEFULL RESTRICTION ENZYME CUT
    STAGGERED WAY- PRODUCING FRAGMENTS WITH STICKY ENDS THAT CAN BOND WITH COMPLEMENTARY STICKY ENDS OF OTHER FRAGMENTS
  10. DNA LIGASE
    ENZYME THAT SEALS THE BONDS BETWEEN RESTRICTION FRAGMENTS
  11. PROCESS
    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
  12. GEL ELECTROPHORESIS
    SEPERATES DNA RESTRICTION FRAGMENTS OF DIFFERENT LENGHTS

    LONGER(SLOWER FRAGMENTS) TOP

    SHORTER(FASTER FRAGMENTS) BOTTOM
  13. RESTRICTION FRAGMENT ANALYSIS
    USEFUL FOR COMPARING TWO DIFFERENT DNA MOLECULES SUCH AS 2 ALLELES FOR A GENE
  14. LIBRARIES
    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
  15. LIBRARIES-FLOW CYTOMETRY
    INVOLVES MARKING THEN SORTING CHROMOSOMES

    SUBGENOMIC FRACTIONS SUCH AS SINGLE CHROMOSOMES CAN BE SORTED USING THIS TECHNIQUE
  16. LIBRARY-CDNA
    CONTAINS COMPLEMENTARY DNA COPIES MADE FROM MRNAS

    REPRESENTS GENES THAT ARE TRASNCRIPTIONALLY ACTIVE AT THE TIME THE CELLS WERE COLLECTED FROM MRNA ISOLATION
  17. CDNA LIBRARY PREPARED BY:
    ISOLATING MRNA FROM CELLS

    • SYNTHESIZING COMP. DNA USING
    • REVERSE TRANSCRIPTASE

    CLONING CDNA MOLECULES INTO VECTOR
  18. LIBRARIES- CLONES CAN BE RECOVERED
    • 2 MAIN TECHNIQS
    • SCREENING

    MAPPING
  19. SCREENING LIBR.
    PROBES COMP. TO PART OF A GENE ARE USED TO SCREEN A LIBR. TO RECOVER CLONES OF A SPECIFIC GENE
  20. SCREENING LIBR PROCESS
    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
  21. LIBRARY MAPPING
    A RESTRICTION MAP ESTABLISHES THE NUMBER AND ORDER OF RESTRCITION SITES AND THE DISTANCE BETWEEN RESTRICTION SITES ON A CLONED DNA SEGMENT
  22. SEQUENCING DNA
    MOST COMMON METHOD OF DNA SEQUENCING IS DIDEOXY CHAIN TERMINATIO SEQUENCING DEVELOPED BY SANGER
  23. SEQUENCING DNA PROCESS
    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
  24. HUMAN GENE THERAPY
    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
  25. DNA FINGERPRINT
    SPECIFIC PATER OF BANDS OF RFLP MARKERS ON A GEL
  26. ANIMAL HUSBANDRY/PHARM ANIMALS
    TRANSGENIC ANIMALS CONTAIN GENES FROM OTHER ORGANISMS
  27. TI PLASMID
    IS THE MOST COMMONLY USED VECTOR FOR INTRUDUCING NEW GENES INTO PLAN CELLS

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