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

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Part 1 PCR:
buffer:30 dNTP:48 glycerol: 60 Formamide: 15
 * lab partner: Crystal Lee
 * Big mix for six reactions (μL):

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

Part 2: PCR modification:
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
 * modify KCl: tube no.1: 10μL(67.5μL water)

done by Crystal Lee
 * modified Mgcl 2 (μL): 0 ; 7.5 ; 20 ; 60

Part3: Viral Growth Curve:
Partners: Crystal, Marla, Anna and above time is according to the room clock; computer time=3:03 the incubation time is all according to the computer time.
 * temperature of water:35 oC
 * Time of phage/host mix: 3:01
 * 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

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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