BIOTECH LAB EXAM 1 CHPT 3

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shockwave
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104217
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BIOTECH LAB EXAM 1 CHPT 3
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2011-09-25 19:26:07
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BIOTECH LAB EXAM CHPTR
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BIOTECH LAB EXAM 1 CHPTR 3
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  1. BACTERIOPHAGE LAMBDA HAS HOW MANY BP IN LENGHT?
    48,502
  2. NAME 3 RESTRICTION ENDONUCLEASE
    • EcoRI
    • BamHI
    • HindIII
  3. NAME 2 BUFFERS USED
    • TRI-BORATE EDTA (TBE)
    • TRIS-EDTA (TE)
  4. T OR F?
    DNA FOR NEAR-TERM USE CAN BE DILUTED WITH DISTILLED OR DEIONIZED WATER?
    TRUE
  5. LOADING DYE IS ------BASED, WHAT HAPPENS WHEN INSERTED IN A WELL?
    SUCROSE. MORE DENSE SINKS INTO WELL.
  6. HOW DO YOU DILUTE DNA?
    • 1. DETERMINE TOTAL VOLUME.
    • (EXPERIMENTS)(TOTAL VOL PER EXPERIMENT)
    • EX:(10 EXPMNTS)(20uL DNA)= 200uL DNA
    • THEN PLUG IN C1V1=C2V2
  7. THE ---- IS THE STANDARD MEASURE OF RESTRICTION ENZYME ZCTIVITY,
    UNIT
  8. DEFINE A UNIT OF RESTRICTION ENZYME ACTIVITY
    THE AMOUNT OF ENZYME NEEDED TO DIGEST TO COMPLETION 1ug OF LAMBDA DNA IN A 50uL REACTION IN 1 HOUR AT 37C.
  9. IN A GEL WHAT IS BUBBLING OFF THE NEGATIVE TERMINAL?
    HYDROGEN
  10. IN A GEL WHAT IS HAPPENING AT THE POSITIVE END?
    ELECTROLYSIS OF WATER--> OXYGEN GAS FORMS BUBBLES.
  11. THE BEST SEPERATION OF ANALYSIS OF LAMBDA AND PLASMID DNA IS ACHIEVED WHEN?
    THE BROMOPHENOL BLUE MIGRATES 40-70 MM FROM THE ORGIN.
  12. THE MIGRATION OF DNA THROUGHT AN AGAROSE GEL DEPENDS UPON WHAT?
    • VOLTAGE.
    • HIGH VOLTAGE YIELDS FASTER MIGRATION.
  13. DEFINE TRANSILLUMINATION
    WHEN LIGHT PASSES THRU THE GEL.
  14. WHY IS A 2X RESTRICTION BUFFER USED INSTEAD OF A 10X BUFFER?
    IT SAVES A PIPETTING STEP TO ADD WATER TO OBTAIN 10 uL VOLUME. EASIER TO ADD 5uL THAN 1uL.
  15. WHAT WAS THE VOLTAGE ON THE BOX IN THE EXPERIEMNT?
    • 100-150 VOLTS
    • (50-100 MILLIAMPS)
  16. T OR F?
    THE ELECTROPHORESE GEL SHOULD ONLY RUN FOR 40-60 MNS OR WHEN BROMOPHENOL BLUE BANDS RUN 40-70mm.
    TRUE
  17. T OR F?
    IT IS NOT A GOOD IDEA TO LET THE DYES RUN OFF THE GEL.
    TRUE
  18. WHY WAS THE WATER ADD TO THE"-" CONTROL?
    To create a negative control. The volume of water added is to maintain the same volume in all of the samples.
  19. WHAT IS THE FUNCTION OF A COMPROMISE RESTRICTION BUFFER?
    A universal buffer that is a compromise between the pH and salt conditions that are preferred by various enzymes.
  20. WHAT IS THE FUNCTION OF A COMPROMISE RESTRICTION BUFFER?
    A universal buffer that is a compromise between the pH and salt conditions that are preferred by various enzymes. PROVIDES Mg++
  21. WHAT ARE THE 2 FUNCTIONS OF THE LOADING DYE?
    • Helps the observer see and monitor the run.
    • Sinks in wells.
  22. WHAT WOULD OCCUR IF THE GEL BOX IS FILLED WITH WATER INSTEAD OF TBE BUFFER?
    SMEARED RUNS
  23. WHAT WOULD OCCUR IF WATER IS USED TO PREPAIR THE GEL INSTEAD OF TBE BUFFER?
    • SMEARED RUNS.
    • THE RESULT WOULD BE POOR SAMPLE ASSIMILATIONS.
  24. WHAT WOULD OCCUR IF THE ELECTRODES ARE REVERSED?
    SHORT RUNS.
  25. WHAT EFFECT WOULD BE OBSERVED IN THE STAINED BANDS OF DNA IN AN AGAROSE GEL IF THE CASTING TRAY IS MOVED WHILE THE GEL IS SOLIDIFYING?
    UNEVEN-WAVE LIKE OR SMEARED.
  26. WHAT EFFECT WOULD BE OBSERVED IN THE STAINED BANDS OF DNA IN AN AGAROSE GEL IF RAN AT HIGH VOLTAGE?
    IT WOULD ACCENTUATE IMPREFECTIONS AND THE GEL WOULD MELT RESULTING IN SMEARED DISTORTED BANDS.
  27. WHAT EFFECT WOULD BE OBSERVED IN THE STAINED BANDS OF DNA IN AN AGAROSE GEL IF A LARGE AIR BUBBLE IS ALLOWED TO SET IN THE GEL?
    DEFORMED OR SMEARED BANDS.
  28. WHAT EFFECT WOULD BE OBSERVED IN THE STAINED BANDS OF DNA IN AN AGAROSE GEL IF TOO MUCH DNA IS LOADED IN A LANE?
    THE DNA WOULD FORM DOUBLETS OR SMEAR.
  29. WHAT EFFECT WOULD BE OBSERVED IN THE STAINED BANDS OF DNA IN AN AGAROSE GEL IF THERE IS INCOMPLETE (PARTICAL) DIGESTION BY A RESTRICTION ENZYME?
    STREAKED OR SMEARED
  30. WHAT CAN BE DONE TO RESOLVE DOUBLET FRAGMENTS?
    • COMPLETEING THE RUN ON A GREATER PERCENT OF AGAROSE GEL.
    • COMPARE GEL % TO SIZE OF DNA.
  31. WHAT IS THE FORMULA TO DETERMINE THE MASS OF DNA IN A GIVEN BAND?
    • (FRAGMENT BP)(CONC. DNA)(VOL DNA)
    • ----------------------------------------------
    • LAMBDA BP

    LAMBDA BP = 48,502
  32. T OR F?
    LAMBDA CAN EXIST AS A CIRCULAR AS WELL AS A LINEAR MODECULE.
    TRUE
  33. AT THE END OF THE LINEAR MOLECULE IS A SINGLE STRANDED SEQUENCE OF 12 NUCLEOTIDES CALLED WHAT?
    • COS SITE
    • AKA STICKY ENDS

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