BIOTEC LAB E1 C7

Card Set Information

Author:
shockwave
ID:
104708
Filename:
BIOTEC LAB E1 C7
Updated:
2011-09-27 17:49:38
Tags:
BIOTEC LAB E1 C7
Folders:

Description:
BIOTEC LAB E1 C7
Show Answers:

Home > Flashcards > Print Preview

The flashcards below were created by user shockwave on FreezingBlue Flashcards. What would you like to do?


  1. IF YOU WERE RUNNING A RAPID AND SENSITIVE ASSEY WHY WOULDN'T YOU USE METHYLENE BLUE?
    DESTAINING USING M. BLUE REQUIRES AT LEAST 30MNS. AND IT COULD FAIL TO DETECT A SMALL BUT POSSIBLE SIGINIFICANT AMOUNT OF UNCUT DNA.
  2. WHAT RESTRICTIVE ENZYMES WERE USED IN LAB 7. WHAT WAS THE REASON?
    • BamHI & HindIII.
    • LIGASED THE KAN OFF ONE PLASMID WHILE OPENING UP A PLACE FOR IT ON THE AMP PLASMID.
  3. IN LAB 7 WHAT WAS THE TYPE OF DNA LIGASE USED AND WHY?
    T4 DNA LIGASE WAS USED. E.COLI DNA LIGASE REQUIRED DIFFERENT REACTION CONDITIONS.
  4. NAME THE 2 UNITS THAT WERE USED TO CALIBRATE LIGASE ACTIVITY.
    • WEISS UNIT
    • COHESIVE-END UNIT
  5. THE PYROPHOSPHATE-EXCHANGE UNIT IS ALSO CALLED WHAT?
    WEISS UNIT
  6. DEFINE THE WEISS UNIT
    • AKA PYROPHOSPHATE-EXCHANGE UNIT.
    • 1 UNIT OF ENZYME CATALYZES THE EXCHANGE OF 1 nMOLE PF 32P FROM PYROPHOSPHATE IN ATP IN 20MNS AT 37C.
    • USED BY IBI & BM.
  7. DEFINE COHESIVE-END UNIT.
    • ONE UNIT OF ENZYME LIGATES 50% OF HindIII FRAGMENTS OF LAMBDA DNA (6ug IN 20uL) IN 30MNS AT 16C
    • USED BY NEB & CBS
  8. 1 WEISS UNIT = ?
    67 COHESIVE UNITS
  9. RESEARCHERS TYPICALLY INCUBATE A LIGATION REACTION HOW LONG & AT WHAT TEMP?
    OVERNIGHT AT ROOM TEMP.
  10. WHAT DNA LIGASE IS INVOLVED WITH HindIII?
    COHESIVE-END UNIT
  11. WHAT DNA LIGASE IS INVOLE WITH ATP ?
    WEISS
  12. BRIEF LIGATIONS OF 2 HRS MINIMUM, IT IS ESSENTIAL TO CHOOSE A HIGH-CONCENTRATION OF T4 DNA LIGASE WITH AT LEAST:
    ------WEISS UNITS/uL OR
    -------COHESIVE-END UNITS/uL.
    • 5 WEISS/uL
    • 300-500 COHESIVE-END UNITS/uL
  13. ------------ BONDING BETWEEN COMPLEMENTARY NUCLEOTIDES ALIGNS BamHI FRAGMENTS.
    HYDROGEN
  14. LIGASE REFORMS -------- BONDS ON EACH SIDE OF THE DNA MOLECULE.
    PHOSPHODIESTER
  15. T OR F?
    WHEN DNA LIGASE MOLECULE IS UTILIZED THEN END PRODUCTS ARE WATER, ADP
    • TRUE.
    • THE ATP IS USED AND REULTS IN ADP. H2O IS H OFF ONE END AND -OH OFF THE OTHER.
  16. T OR F?
    LIGATION OF THE FOUR BamHI/HindIII RESTRICTION FRAGMENTS OF pAMP AND pKAN PRODUCES MANY TYPES OF HYBRID MOLECULES INCLUDING PLASMIDS POSSESING MORE THAN 2 FRAGMENTS.
    TRUE
  17. THE COMPLETE pAMP HAS HOW MANY BP?
    4539
  18. AFTER BamHI & HindIII DO THEIR THING TO pAMP, WHAT ARE THE 2 BP SIZES?
    • 3755 (CONATINS AMP)
    • 784 (SIDE PIECE)
  19. HOW MANY BP IN A COMPLETE pKAN PLASMID?
    4207
  20. AFTER BamHI & HindIII CUT UP pKAN, WHAT ARE THE 2 BP SIZES?
    • 1861 (CONTAINS kan)
    • 2332 (CONTAINS ORI)
  21. 3 DIFFERENT REPLICATING PLASMIDS ARE CREATING BY LIGATING TWO BamHI/HindIII FRAGMENTS, NAME THEM.
    • LIGATION OF 784 & 3755 REGENERATES pAMP
    • LIGATION OF 1861 & 2332 REFORMS pKAN
    • LIGATION OF 1861 & 3755 PRODUCE THE "SIMPLE RECOMBINATE" PLASMID pAK. THE KANAMYSIN-RESISTANT GENE FUSED TO pAMP BACKBONE.
  22. WHY IS ATP ESSENTIAL FOR THE LIGASE REACTION.
    UTILIZATION OF THE PHOSPHEROUS AND THE ENERGY IN THE BOND.
  23. WHAT THE FUCK ARE STICKY ENDS?
    • The ends of the cut have an overhanging piece of single-stranded DNA.
    • These are called "sticky ends" because they are able to base pair with any DNA molecule containing the complementary sticky end.
    • In this case, both DNA preparations have complementary sticky ends and thus can pair with each other when mixed.
    • a DNA ligase covalently links the two into a molecule of recombinant DNA.

What would you like to do?

Home > Flashcards > Print Preview