E. Strand, E. Chille 20190528 DNA/RNA Extractions, Qubit
E. Strand 20190529 Tape Station
E. Strand, M. Schedl 20190529 Ship Samples

20190529 Tape Station, Ship Samples

Protocol: TapeStation

TapeStation Notes:

• Tape and buffer out at 9:18 am, done at 9:48 am
• Thermocycler start 10:09 end 10:15
• TapeStation start 10:19 end 10:29
• TapeStation did not read last sample (20, 221, C2) because I aborted the run right after I clicked start so I could spin the samples down, then started the TapeStation again and didn’t realize the Tape had already been scanned once. Instead of 10 samples left, there were 9 and the 10th sample in our extractions wasn’t read. The samples had not yet been pierced when I aborted the first run.

TapeStation Results:

Extraction #	Sample ID	Coral ID	Content	TapeStation ID	RIN^e
11	784	TBR7_52	Eggs	B1	4.1
12	215	TBR7_55	Eggs	C1	3.8
13	760	FR3_61	Eggs	D1	3.8
14	790	TBR7_50	Sperm	E1	2.2
15	19	FR3_62	Eggs	F1	3.6
16	185	FR3_64	Sperm	G1	3.1
17	173	FR3_63	Sperm	H1	1.8
18	754	FR3_61	Sperm	A2	1.3
19	167	TBR7_53	Eggs	B2	4.3
20	221	FR3_60	Sperm	C2	NA

We want 500 ng of 5 TBR7 eggs and 5 FR eggs that have a matching sperm with DNA content. Below are the samples we decided to ship:

Extraction #	Sample ID	Content	Coral ID	Eggs RNA (ug)	Sperm DNA (ug)
11	784	eggs	TBR7_52	9.18	8.12
12	215	eggs	TBR7_55	9.6	2.86
19	167	eggs	TBR7_53	11.5	4.54
1	796	eggs	TBR7_50	4.25	5.77
4	203	eggs	TBR7_54	6.11	0
13	760	eggs	FR3_61	10.36	5.27
15	19	eggs	FR3_62	14.54	2.98
3	227	eggs	FR3_60	17.01	13.07
6	179	eggs	FR3_63	2.51	8.68
7	19	eggs	FR3_64	7.03	7.51

• TBR7_54’s sperm sample did not have any DNA, but we shipped the eggs sample anyway so we could have 5 TBR and 5 FR samples.

Aliquoting the samples: M. Schedl aliquoted extraction numbers 1, 3, 4, 6, and 7 previously.

Acropora ID	ng/ul (Qubit avg)	sample volume	water volume	concentration (ng/uL)
784	96.6	5.18	4.82	50
215	101	4.95	5.05	50
167	121	4.13	5.87	50
760	109	4.59	5.41	50
19	153	3.27	6.73	50
796	44.7	11.19	NA	44.7
203	64.3	7.78	2.22	50
227	179	2.79	7.21	50
179	26.4	18.94	NA	26.4
191	74	6.76	3.24	50

• ng/uL is the average reading from the Qubit
• sample volume is ng/uL value divided by 500 to get how much volume of sample is needed to have 500 ng of RNA
• water volume is how much volume of water is needed to have at least 10 uL of liquid to ship
• concentration is how many ug of sample per uL is in the shipment

Other information for shipping:

• Species: Acropora hyacinthus
• Sample Buffer: ultra pure water
• Purification Method: Zymo-Duet RNA DNA Extraction Kit
• Sample Type: Total RNA