CaCl2 Transfection

Date:
Experiment:
Aim of expt:
Samples:

Conditions:
1. DNA
2. CaCl2
3. 2XBBS
4. Treatment
5. Harvest
Procedure:
1. Split _______________ on ______________. Cell count =
2. Incubate until 50-70% confluent. Time =
3. Transfect on _______________
3.5cm dish

6cm dish

10cm dish

15cm dish

DNA

4g total

10g total

22ug total

50g total

1X CaCl2

100 l

237 l

500 l

1 ml

2X BBS

100 l

237 l

500 l

1 ml

Plates

DNAs

1. Prepare DNA in 1.5ml microtube:

Mix by vortex and quick spin down
2. Add 1X CaCl2
Diluted 10X CaCl2 to 1X with sterile water immediately before use.
Vortex and spin down.
3. Add 2X BBS (pH 6.95)
Add drop by drop, mix by light tap, grading increase BBS concentration.
Finally use pipette mix couple times.
4. Incubate at RT for 15 min.
5. Add DNA drop by drop equally into cell plate (50~70% confluence), mix well and gently.
4. Incubate at 37C, 5% CO2, _____________ hrs
5. Observe under UV-Microscope before changing media.
Observations:

6. Change media on ______________

7. Observe under UV-Microscope before harvesting cells.
Observations:

8. Harvest cells on ______________

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