LABCRISPR
EXPERIMENT GOAL: we generate bacteria cells in PLATE 1 – cells that cannot survive in the harsh environment of PLATE 2. Then we apply CRISPR and edit the gene such that the bacteria cells can survive in PLATE 2.
STEP4_0_10
STEP4_0_8
CRISPR KIT
SEQUENCE
01
PROCEDURE Bacteria transformation 100μL in empty microcentrifuge
2:30pm
CHECK
STEP4_1_6
TIME
SEQUENCE
02
PROCEDURE Mixing bacteria
2:30pm
CHECK
Step4_2_4
STEP4_2_2
TIME
SEQUENCE
03
PROCEDURE Mixing Cas9, gRNA, rpss w/ bacteria (※ Must do in order)
STEP4_3_2
STEP4_3_2
TIME
2:30pm
CHECK
SEQUENCE
04
TIME
PROCEDURE
Placed in fridge 0-4degrees Celsius for 30 minutes
SEQUENCE
05
2:30pm
CHECK
PROCEDURE Place in hot water (42-45 Celsius) for 30 seconds
3:00pm
CHECK
STEP4_5_6
STEP4_5_1
TIME
SEQUENCE
06
PROCEDURE Mix LB Media w/ 1.5ml of water and stir gently
STEP4_3_2
STEP4_6_2
TIME
3:00pm
CHECK
SEQUENCE
07
PROCEDURE Repeat (6) and (5) 100μL 5 times
3:00pm
CHECK
STEP4_1_6
TIME
08
PROCEDURE After 4-12 hours, placed bacteria in plate 2
STEP4_8_1
SEQUENCE
09
SEQUENCE
4:05pm
CHECK
PROCEDURE Place cover on plate 2 and wait 24-48 hours checking for results
STEP4_9_1_15 hours later
SEQUENCE
TIME
NEXT DAY STEP4_9_2_24 hours later
9:00pm
10
PROCEDURE Success!
CHECK
TIME
9:10pm
CHECK