User:Mark X. Ling/Notebook/Micb 323/2009/01/13

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Experiment 1 (Jan 13, 2009)

Part 1 PCR:

  • lab partner: Crystal Lee
  • Big mix for six reactions (μL):
 buffer:30
 dNTP:48
 glycerol: 60
 Formamide: 15
  • PCR done:
    • 1: Primer set 1 +DNA 1
    • 2: Primer set 1 +DNA 2
    • 3: Primer set 2 +DNA 1
    • 4: Primer set 2 +DNA 2
    • 5: Primer set 1 +no DNA (control)

Primer set 1 : otrA1 and otrA3 Primer set 2: otrA1 and otrA2

Part 2: PCR modification:

  • modify KCl: tube no.1: 10μL(67.5μL water)

no.2: 0μL (77.5μL water)
no.3: 20μL (57.5μL water)
no.4: 70μL (7.5μL water)
rest: tris: 10μL, MgCl2:7.5μL, tween: 5μL

  • modified Mgcl 2 (μL): 0 ; 7.5 ; 20 ; 60

done by Crystal Lee

Part3: Viral Growth Curve:

Partners: Crystal, Marla, Anna and

  • temperature of water:35 oC
  • Time of phage/host mix: 3:01

above time is according to the room clock; computer time=3:03

  • T0 = 3:08; T1=3:13; T2=3:18; T3=3:23; T4=3:28; T5=3:38; T6=3:48; T7=3:58; T8=4:08; T9=4:18

the incubation time is all according to the computer time.

Post Lab comments

  • PCR preparation went smoothly.
  • Temperature of water was slightly lower than one specified in manual.
  • overall the procedures in this lab was conducted according to the manual and went smoothly


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