Lab Crispr Presentation

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