Project: Porites Bleaching 2019

Goal

Dilute extracted RNA to 10 ug/uL in 50 uL and consolidate into one plate for Tag-seq. Molecular QC information can be found here.

Samples

Fragment ID	Timepoint	Group	Vial #	Extraction Date	RNA Conc (ng/uL)	Input for 10ng/ul (uL)	Dilutent Vol (uL)	Total Vol (uL)	Well
R29	A1	Bleached	44	20201117	13.2	37.88	12.12	50	A1
R20	A4	Bleached	23	20201106	15.6	32.05	17.95	50	A2
R19	A4	Bleached	49	20201119	21.8	22.94	27.06	50	A3
R7	A2	Control	37	20201117	24	20.83	29.17	50	A4
R7	A4	Control	34	20201111	25.4	19.69	30.31	50	A5
R40	A1	Control	88	20201202	33.8	14.79	35.21	50	A6
R37	A4	Bleached	74	20201125	19.2	26.04	23.96	50	A7
R26	A4	Mortality	87	20201202	13.2	37.88	12.12	50	A8
R29	A4	Bleached	65	20201120	15.6	32.05	17.95	50	A9
R26	A1	Mortality	46	20201117	16.4	30.49	19.51	50	A10
R40	A2	Control	14	20201029	18.4	27.17	22.83	50	A11
R19	A2	Bleached	39	20201117	14.8	33.78	16.22	50	A12
R28	A4	Mortality	24	20201106	28	17.86	32.14	50	B1
R35	A2	Mortality	13	20201029	15.2	32.89	17.11	50	B2
R28	A2	Mortality	36	20201111	10.2	49.02	0.98	50	B3
R36	A4	Mortality	40	20201117	14.2	35.21	14.79	50	B4
R20	A1	Bleached	30	20201110	25.6	19.53	30.47	50	B5
R17	A1	Control	15	20201029	18.8	26.6	23.4	50	B6
R40	A4	Control	80	20201126	20.4	24.51	25.49	50	B7
R32	A4	Control	33	20201111	13.4	37.31	12.69	50	B8
R37	A2	Bleached	53	20201119	12.6	39.68	10.32	50	B9
R28	A1	Mortality	71	20201125	20.6	24.27	25.73	50	B10
R11	A2	Mortality	22	20201106	31.4	15.92	34.08	50	B11
R8	A2	Bleached	58	20201120	14.2	35.21	14.79	50	B12
R17	A2	Control	51	20201119	14	35.71	14.29	50	C1
R11	A4	Mortality	63	20201120	20.8	24.04	25.96	50	C2
R11	A1	Mortality	32	20201111	38.8	12.89	37.11	50	C3
R23	A2	Control	11	20201027	15.8	31.65	18.35	50	C4
R7	A1	Control	56	20201119	25.2	19.84	30.16	50	C5
R35	A4	Mortality	41	20201117	10.8	46.3	3.7	50	C6
R23	A4	Control	73	20201125	13.4	37.31	12.69	50	C7
R32	A2	Control	35	20201111	17.8	28.09	21.91	50	C8
R8	A4	Bleached	43	20201117	25	20	30	50	C9
R23	A1	Control	67	20201125	15	33.33	16.67	50	C10
R32	A1	Control	54	20201119	27	18.52	31.48	50	C11
R29	A2	Bleached	21	20201106	29.2	17.12	32.88	50	C12
R17	A4	Control	89	20201202	25.4	19.69	30.31	50	D1
R37	A1	Bleached	10	20201027	24.6	20.33	29.67	50	D2
R35	A1	Mortality	68	20201125	15.8	31.65	18.35	50	D3
R8	A1	Bleached	31	20201111	13	38.46	11.54	50	D4
R20	A2	Bleached	12	20201027	27.4	18.25	31.75	50	D5
R26	A2	Mortality	57	20201120	23.2	21.55	28.45	50	D6
R36	A2	Mortality	62	20201120	12.4	40.32	9.68	50	D7
R36	A1	Mortality	55	20201119	16.2	30.86	19.14	50	D8
R19	A1	Bleached	29	20201110	36.4	13.74	36.26	50	D9

Plate Map

Protocol

1) Remove samples from the -80°C freezer and place on a tube rack in an ice bucket to thaw.

2) Fill each well of the 96-well PCR plate with the corresponding diluent volume (I used Ultra Pure Water).

3) Place plate onto a block frozen at -80°C.

4) Once samples are thawed, vortex and spin down the sample before adding the appropriate volume into the corresponding well.

5) Seal plate with aluminum foil tape and seal each well. Write on the label and the side of the plate with the following information:

• Date
• Name
• Project
• Job Number

6) Place remaining samples and plate back into the -80°C freezer.