SOE PCR for clpX

2/23/2015

3 PCR 1.5 μL (PCR product from clpX 3)
5 PCR 1.5  μL (PCR product from clpX 5)
clpX3 R primer 1.5 μL
clpX5 F primer 1.5 μL
supermix 21 μL


Protocol:
 95 °C    3 minutes

95 °C   30 seconds
55 °C  2 minutes                    30 cycles
68 °C  1.5 minutes

Final elongation 68 °C  for 5 minutes.

1% agarose run for 4 hours.

PCR for LVS chiT3, clpX3,, clpB5 and chiT5

2/11/2015

Protocol 5

95 c                    3 minutes

95 c                    30 second

44 c                     1.5 minutes                                35 cycle

68 c                    1.5 minute
Final elongation 5 minutes, total volume 25 microliter.

Samples:
1-chiT3
2-clpX3
3- clpB5
4
- chiT5
1.5 mico primares x 2
2 micro g DNA
20 micro supermix.

Run on 1% gel for 3 hours.

PCR for LVS clpB3, chiT3, clpX5, clpB5 and chiT5

2/8/2015

Protocol 4

95 c                    3 minutes

95 c                    30 second

46 c                     30 seconds                                  35 cycle

68 c                    1 minute

Final elongation 5 minutes, total volume 25 microliter.

Samples:
1- clpB3
2-chiT3
3-clpX5
4- clpB5
5- chiT5
1.5 mico primares x 2
2 micro g DNA
20 micro supermix.
Run on 1% gel for 3 hours.

PCR for LVS clpX3, clpB5 and chiT5

2/2/2015

Protocol 3

95 c                    7 minutes

95 c                    30 second

49 c                     30 seconds                                  35 cycle

68 c                    1 minute

Final elongation 5 minutes, total volume 25 microliter.

Samples:1- clpX 3

1- clpX 3

2-clpB 5

3-chiT 5

Result:  clpX 3 showed band

Run on 1% gel for 3 hours.