RNA isolation

Isolating RNA from the Col-0 infiltrations yesterday for the AtGRP7 stuff. Also getting the UPF RNA without any treatment, just to see if PR-1 is constitutively on compared to Col-0.

Samples:
1        Col-0
2        UPF1-4
3        UPF1-5
4        Mock 5hpi
5        P1 5hpi
6        P5 5hpi
7        Mock 7hpi
8        P1 7hpi
9        P5 7hpi
10        Mock 12hpi
11        P1 12hpi
12        P5 12hpi

3 chloroform washes. Ended up with 500ul of sample, added 500ul isopropanol and put in the -20.

12/22/10 12:17 PM
Spinning for 10 minutes at room temperature.

12/22/10 1:01 PM
Made mastermix for DNAse:
1 glycogen
3 10x DNAse buffer
26 DEPC H2O

Resuspended the pellets in 29ul of this mastermix, and then added 1ul DNAse. 37* for 10 minutes, followed by 70* for 10 minutes. Added 75 100% EtOH, then -20*.

12/22/10 4:05 PM
Spinning for 10 minutes at room temperature.

40ul resuspension in DEPC water.

Sample ID	ng/ul	260/280	260/230
Col-0	322.73	2.09	2.29
UPF1-4	230.38	2.09	2.27
UPF1-5	264.25	2.1	2.29
Mock 5hpi	368.82	2.09	2.25
P1 5hpi	415.91	2.09	2.25
P5 5hpi	447.06	2.1	2.27
Mock 7hpi	324.73	2.09	2.27
P1 7hpi	231.85	2.1	2.27
P5 7hpi	380.32	2.1	2.26
Mock 12hpi	375.76	2.08	2.23
P1 12hpi	431.2	2.09	2.26
P5 12hpi	305.23	2.09	2.21

The CGRB website isn’t working, so I will submit this for bioanalyzer tomorrow.