Processing the E5 January, March, September, and November 2020 Instantaneous Calcification Samples

Goal

Process calcification samples collected for the E5 uROL Epigenetics Project from January, March, September, and November 2020 (timepoints 1, 2, 3, and 4). The samples were collected in January (n = 146), March (n = 151), September 2020 (n = 114), and November 2020 (n = 145) and fixed with 75uL of saturated mercuric chloride for transportation back to URI on November, 22nd 2020. All samples were brought back from the January and March timepoints November 2020, but only 51 bottles could fit in the containers from the September timepoint. The remaining samples for the September and November 2020 timepoints are as follows: the Silbiger lab brought back the remaining samples from Mo’orea for the September 2020 timepoint in August 2021 and processed them on their titrator in October 2021 following the Silbiger Lab Titrator Protocols. The November 2020 samples were brought back to URI by Hollie and Ariana in November 2021. All remaining samples are in the URI CBLS 180 Environmental Room.

Samples Remaining

• January 2020, 146 samples remaining
• September 2020, 51 samples remaining
• November 2020, 145 samples remaining

Process

Samples were collected, experimental run, and transported back to URI following the E5 Instantaneous Calcification Protocol in Mo’orea, French Polynesia.

Upon the return to URI, we wanted to test the total alkalinity signal we would receive from our initial seawater samples before the experimental procedure and after to calculate delta total alkalinity. To do so, titrations will need to be completed for all samples to calculate total alkalinity following the alkalinity anomaly technique (Chisholm and Gattuso, 1991).

All samples will be processed following the established Putnam Lab Titrator Protocol.

All data processed and collected on the Putnam Lab Titrator will be available on the Putnam Lab Titrator GitHub Repository.

Titration Data Entry Process

• After uploading titration data to the Putnam Lab Titrator Repository from the CBLS lab computer, you will need to enter this into the E5 uROL Epigenetics Project Timeseries Data.
• First, copy the titrator TA outputs for each sample into the respective TP1, TP2, TP3, and TP4 data sheets found in the E5 uROL Epigenetics Project Timeseries Data Page.

You can find the information highlighted in green from the labels on the bottles, and the rest of the information can be found using the instructions below

• Second, you will need to match the corresponding Mo’orea measurements (i.e. lab pH, volume, salinity, and temperature in Mo’orea) and information into these data sheets as well.
  • The information for this can be found in each individual timeseries 1, 2, 3, and 4 folders on the E5 uROL Epigenetics Project Timeseries Data Page.
  • To find this corresponding information for specific samples and blanks, you will look at the 1_DeltaTA_metadata.csv, 2_DeltaTA_metadata.csv, 3_DeltaTA_metadata.csv, and 4_DeltaTA_metadata.csv data sheets.
  • To find this information for the initial water samples, you will look at the 2_initial_TA_samples.csv, 3_initial_TA_samples.csv, and 4_initial_TA_samples.csv data sheets. I have not been able to find the initial sample data sheet for TP1 so you may need to ask Ariana and/or Hollie.
• I have completed all of the samples for timepoint 2 (March 2020) so if you want to see data examples for comparison that is a good place to look.

Data and notes for testing pH probe and buffers on 20210221

Notes

• In the initial titration set-up I noticed that the pH calibration was a bit off due to the zero point and slope values we received here. I ran a CRM and found that is was within the < 1% accuracy that we would expect but at a value of -0.82% was a little high especially with the floating pH calibration data. I began to troubleshoot the problem and thought that maybe the pH buffers could be off from prior knowledge. A new round of buffers was ordered to test the pH probe. pH calibration and CRM accuracy information can be found in the Putnam Lab Titrator Protocol.

Data

SampleID	TA	Mass	Salinity
JUNK1	1791.44566335087	59.81	35
CRM 1	2215.55476501538	59.561	33.417

Data and notes for testing variability in CRMs and pH probe 20210228

Notes

• Continued the troubleshooting process and still saw that the pH calibration values were off and the probe was floating in its mV values when measured manually for 40 seconds in each of the 4, 7, and 10 pH buffers. The values were small but still seemed inconsistent. To test how much variability we were seeing in CRMs due to the pH probe, I ran 8 CRMs. There was a lot of variability between the CRMs so a new pH probe was ordered and received on 20210304.

pH buffer 4 manually measured values (mV)

pH buffer 7 manually measured values (mV)

pH buffer 10 manually measured values (mV)

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210218	2215.547681	2234.07	-0.829084093	141	CRM opened 20210216
20210221	2215.554	2234.07	-0.828801246	141	CRM opened 20210221
20210228	2211.139264	2234.07	-1.026410826	141	CRM opened 20210216
20210228	2213.599263	2234.07	-0.916297939	141	CRM opened 20210216
20210228	2218.811032	2234.07	-0.683012077	141	CRM opened 20210228
20210228	2208.830425	2234.07	-1.129757569	141	CRM opened 20210228
20210228	2203.497378	2234.07	-1.368472003	141	CRM opened 20210228
20210228	2213.354976	2234.07	-0.927232556	141	CRM opened 20210228
20210228	2230.94523	2234.07	-0.13986896	141	CRM opened 20210228
20210228	2222.545765	2234.07	-0.515840388	141	CRM opened 20210228

Total Alkalinity Output

SampleID	TA	Mass	Salinity
JUNK1	932.4357056	59.701	35
CRM1	2211.139264	59.861	33.417
CRM2	2213.599263	60.436	33.417
CRM3	2218.811032	60.178	33.417
CRM4	2208.830425	60.307	33.417
CRM5	2203.497378	60.315	33.417
CRM6	2213.354976	60.201	33.417
CRM7	2230.94523	60.413	33.417
CRM8	2222.545765	60.105	33.417

New pH probe received on 20210303

New pH probe assembled, installed and tested on 20210307

Data and notes for testing variability in CRMs and new pH probe 20210314

Notes

• Continued the troubleshooting process and had great pH calibration values with the new probe and buffers before any runs today. Purged all bubbles and checked every box before running one junk and 8 CRM samples. After the run with 8 CRMS, I noticed that the variability from the CRM value was still >1% in a few of the samples. I attributed this to the older CRM bottle that was opened on 20210228. I opened a brand new CRM bottle and ran four samples but there was still variability to the original CRM value but less than in the previous run. I called Mettler Toledo support and they said this could be an offset with the titrant (acid) and they have had this happen. We could measure the titrant against tris buffer to calculate the offset and apply that to our equations. Noticed that the titrant was replaced in May 2019, which seemed very old and think evaporation could have effected its composition. Next round I am going to change out the titrant with a new bottle.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210314	2182.69197	2234.07	-2.29975022	141	CRM opened 20210228
20210314	2183.070705	2234.07	-2.282797543	141	CRM opened 20210228
20210314	2179.989317	2234.07	-2.420724647	141	CRM opened 20210228
20210314	2180.118916	2234.07	-2.414923637	141	CRM opened 20210228
20210314	2194.262889	2234.07	-1.781820203	141	CRM opened 20210228
20210314	2194.808799	2234.07	-1.757384566	141	CRM opened 20210228
20210314	2192.134088	2234.07	-1.877108222	141	CRM opened 20210228
20210314	2192.253611	2234.07	-1.871758246	141	CRM opened 20210228
20210314	2205.731211	2224.47	-0.842393439	180	CRM opened 20210314
20210314	2194.005521	2224.47	-1.369516288	180	CRM opened 20210314
20210314	2192.044753	2224.47	-1.457661693	180	CRM opened 20210314
20210314	2196.378938	2224.47	-1.262820467	180	CRM opened 20210314

Total Alkalinity Output

SampleID	TA	Mass	Salinity
JUNK1	1981.003448	60.113	35
CRM1	2182.69197	60.225	33.417
CRM2	2183.070705	59.774	33.417
CRM3	2179.989317	60.107	33.417
CRM4	2180.118916	60.161	33.417
CRM5	2194.262889	59.659	33.417
CRM6	2194.808799	59.719	33.417
CRM7	2192.134088	59.651	33.417
CRM8	2192.253611	59.892	33.417
CRM1	2205.731211	60.297	33.623
CRM2	2194.005521	60.319	33.623
CRM3	2192.044753	59.881	33.623
CRM4	2196.378938	59.971	33.623

Data and notes for testing variability in CRMs with new titrant 20210328

Notes

• Continued the troubleshooting process and changed out the older titrant with a new Batch #A16 titrant. Purged the lines four times to make sure to run through any of the older titrant from the lines. Ran a pH calibration that had sound numbers and then I ran a junk sample before five CRMs. IT WORKED! The CRM values were very accurate and everything looked good from the run. Can continue with samples moving forward.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210328	2766.991908	2225.47	24.33292331	180	CRM opened 20210314
20210328	2226.220045	2226.47	-0.011226509	180	CRM opened 20210314
20210328	2224.425627	2227.47	-0.136674041	180	CRM opened 20210314
20210328	2224.201971	2228.47	-0.191522852	180	CRM opened 20210314
20210328	2225.549637	2229.47	-0.175842846	180	CRM opened 20210314
20210328	2228.292488	2230.47	-0.097625722	180	CRM opened 20210314

Total Alkalinity Output

SampleID	TA	Mass	Salinity
JUNK1	2766.9919084924	60.137	35
CRM1	2226.22004515256	60.18	33.623
CRM2	2224.4256267384	60.274	33.623
CRM3	2224.20197069824	60.29	33.623
CRM4	2225.54963650217	60.077	33.623
CRM5	2228.2924875591	60.135	33.623

Data and notes for testing E5 samples TA signal 20210504

Notes

• The CRM values were very accurate and everything looked good from the previous run, testing a batch of samples from March (TP2) run 1 on 20200304 to see if there is a processable deltaTA signal from the initial sample to the blank and other coral sample chambers. All went well and we were able to see > 30 for deltaTA in almost all samples which is a good value moving forward. Going to run more samples from another run to make sure this occurs in other replicates.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210504	2224.823	2224.47	0.015869	180	CRM opened 20210418

Total Alkalinity Output

SampleID	TA	Mass	Salinity
JUNK1	2030.80445223374	60.035	35
INITIAL-2	2325.79665116253	59.525	36.03
ACR-225	2229.92073279084	60.488	36.03
POC-215	2271.34957005683	59.695	36.02
BLANK-1	2298.63819200678	60.297	36.03
POR-224	2206.22121318086	60.266	36

Data and notes for testing E5 samples TA signal 20210506

Notes

• The CRM values were very accurate and everything looked good from the previous run, testing a batch of samples from March (TP2) run 2 on 20200304 to see if the deltaTA values are also representative of what we saw in yesterdays runs. Most of the samples again had deltaTA’s > 30 and all looked good.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210506	2225.094	2224.47	0.028052	180	CRM opened 20210418

Total Alkalinity Output

SampleID	TA	Mass	Salinity
JUNK1	2018.39880477983	60.226	35
INITIAL-1	2309.66768581353	60.083	37.38
INITIAL-2	2310.6341993138	59.885	37.38
BLANK-2	2312.66284694853	59.917	37.4
POC-201	2302.40007114124	59.624	37.4
ACR-244	2277.31062313817	59.786	37.39
POR-235	2288.18858541504	59.525	37.41
ACR-229	2272.13489556652	59.84	37.38
POR-245	2268.37232119263	60.317	37.43

Data and notes for processing E5 samples TP2, Run 5 20210511

Notes

• Had to completed two seperate titration runs. All samples looked good. Updated timeseries data to urol-e5 GitHub.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210511	2228.879	2224.47	0.198204516	180	CRM opened 20210418

Total Alkalinity Output - Titration Run 1 (8 samples)

SampleID	TA	Mass	Salinity
JUNK1	2023.62860758224	60.323	35
ACR-360	2299.24809778777	59.666	37.3
POR-355	2272.98399534722	60.365	37.29
POC-394	2331.64157338251	59.807	37.3
ACR-351	2308.62561873151	59.796	37.27
BLANK-5	2341.16654731596	59.832	37.3
INITIAL-1	2349.40134517588	60.137	37.3
ACR-368	2335.36944848248	60.295	37.32
POR-349	2312.55677108142	59.746	37.28

Total Alkalinity Output - Titration Run 2 (4 samples)

SampleID	TA	Mass	Salinity
JUNK1	2020.61803398065	60.11	35
POR-367	2271.59356605544	59.959	37.32
POR-353	2246.61383181666	60.357	37.32
POC-359	2320.53306339006	59.601	37.29
INITIAL-2	2354.906625	59.539	36.52

Data and notes for processing E5 samples TP2, remaining Run 1 samples and 1 run 5 sample 20210525

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210525	2223.505163	2224.47	-0.043373783	180	CRM opened 20210418

Total Alkalinity Output

SampleID	TA	Mass	Salinity
JUNK1	2021.113958	59.52	35
INITIAL-1	2328.815162	59.78	36.1
POR-214	2166.262087	59.875	36.12
POC-200	2257.931561	60.457	36.08
POC-255	2290.17382	60.446	36.1
ACR-237	2282.117545	59.516	36.11
POR-253	2121.493182	59.948	36.09
ACR-258	2263.09945	59.723	36.12
INITIAL-2	2354.906625	59.539	36.52

Data and notes for processing E5 samples TP2, remaining Run 2 samples (4), 9 samples from run 3 20210526

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub. Started to record the temperature of seawater in the room for each day. Today it was 24.8°C.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210526	2223.47	2224.47	-0.044954528	180	CRM opened 20210418

Total Alkalinity Output

SampleID	TA	Mass	Salinity	Run
POC-257	2294.21281589112	60.182	36.79	2
POR-240	2253.77127801894	60.261	36.88	2
POR-262	2248.02673011785	59.802	36.87	2
POC-254	2287.41202551437	60.339	36.86	2
ACR-265	2283.76004008528	59.977	37	3
POC-239	2295.81472407995	60.457	37.03	3
INITIAL-1	2315.48416509178	59.894	37.02	3
POR-209	2254.08211275199	60.23	37.02	3
POC-217	2299.82917108649	60.113	37.01	3
BLANK-3	2308.18866510965	59.689	37.02	3
POR-221	2172.68917435377	60.417	37.01	3
POC-259	2285.3899644917	59.997	37.04	3
POC-207	2299.9880480591	59.653	37.04	3

Data and notes for processing E5 samples TP2, remaining Run 3 samples (3), 5 samples from run 4 20210527

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub. Temperature of room seawater: 24.5°C.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210527	2221.347	2224.47	-0.140392992	180	CRM opened 20210418

Total Alkalinity Output

SampleID	TA	Mass	Salinity	Run
JUNK1	2048.4918806143	60.049	35	-
POR-236	2210.78794910016	59.592	35.35	3
POR-206	2169.18082109917	60.638	35.38	3
INITIAL-2	2314.88140617624	60.075	35.35	3
POR-260	2215.42467558492	59.651	35.5	4
POC-222	2294.11805543367	59.754	35.49	4
POR-242	2002.10749874344	60.135	35.5	4
POR-266	2128.69485050727	60.084	35.49	4
POC-248	2291.29416890313	60.283	35.5	4

Data and notes for processing E5 samples TP2, remaining Run 4 samples (7), 9 samples from run 6 20210601

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub. Temperature of room seawater: 24.2°C.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210601	2222.281343	2224.47	-0.098390052	180	CRM opened 20210526

Total Alkalinity Output

SampleID	TA	Mass	Salinity	Run
INITIAL-2	2323.47645891917	60.309	35.66	4
BLANK-4	2323.38824894761	60.327	35.64	4
POC-219	2314.63393459143	60.164	35.64	4
POR-216	2162.56151837602	59.636	35.64	4
INITIAL-1	2302.12915601656	59.576	35.66	4
POC-205	2306.82987290008	60.252	35.64	4
POR-383	2292.09784313125	59.837	35.83	6
POR-381	2268.80549000996	60.483	35.84	6
POR-341	2303.485165	59.517	35.82	6
POC-238	2290.323051	59.502	35.82	6
BLANK-6	2330.885054	59.868	35.81	6
POC-378	2317.066965	60.182	35.83	6
POC-366	2319.944227	59.805	35.8	6
INITIAL-1	2335.442874	59.799	35.84	6
ACR-396	2313.47919	59.592	35.85	6
INITIAL-2	2369.881136	59.78	35.82	6

Data and notes for processing E5 samples TP2, remaining Run 6 samples (3), 12 samples from run 7 20210624

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub. Added temperature probe to titrator, continuous temperature being measured during runs.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210624	2228.452	2224.47	0.179008932	180	CRM opened 20210526

Total Alkalinity Output

SampleID	TA	Mass	Salinity	Run
JUNK1	2083.83581489909	59.875	35	NA
ACR-350	2313.45289533546	59.604	37.56	6
ACR-343	2312.10286349871	60.376	37.58	6
POC-391	2308.18425528784	60.361	37.7	7
BK-7	2329.54336354651	59.908	37.66	7
POC-377	2318.93041886119	59.958	37.66	7
ACR-347	2287.77157182072	60.403	37.66	7
POR-357	2301.85458181472	59.963	37.61	7
POR-384	2068.74678641006	60.493	37.61	7
POC-371	2333.22335273167	59.653	37.65	7
POC-373	2319.57500582734	59.598	37.61	7
INITIAL-1	2332.42150299334	59.664	37.68	7
POR-338	2296.40931234685	60.389	37.66	7
ACR-364	2313.9874371687	59.822	37.65	7
INITIAL-2	2331.80262384803	59.961	37.66	7

Data and notes for processing E5 samples TP2, Run 8 samples (12), 1 samples from run 9 20210706

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210706	2230.091863	2224.47	0.25272822	180	CRM opened 20210526

Total Alkalinity Output

SampleID	TA	Mass	Salinity	Run
JUNK1	2127.87573977338	59.736	35	NA
POR-365	2302.22568628682	60.488	37.66	8
POC-369	2332.48754752583	59.741	37.64	8
INITIAL-2	2338.97139177075	59.892	37.61	8
POR-340	2267.76065570345	59.596	37.64	8
BLANK-8	2334.95502705341	60.035	37.63	8
POR-385	2268.85278894355	59.742	37.65	8
POC-372	2321.65688763208	59.923	37.68	8
POC-375	2303.3815103305	60.203	37.65	8
POC-346	2325.31511305514	59.651	37.77	8
ACR-393	2319.92989417533	60.172	37.72	8
INITIAL-1	2336.42991421746	60.005	37.74	8
POC-395	2316.5050298392	59.603	37.74	8
POC-48	2325.01226734367	59.609	37.75	9

Data and notes for processing E5 samples TP2, Run 9 samples (8) 20210708

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210708	2228.957083	2224.47	0.201714718	180	CRM opened 20210526

Total Alkalinity Output

SampleID	TA	Mass	Salinity	Run
JUNK1	2265.60200372629	59.841	35	NA
POC-42	2322.61351513766	60.339	37.88	9
POR-73	2216.31310048821	60.323	37.96	9
ACR-186	2294.35842028192	60.143	37.95	9
BLANK-9	2340.47696082826	60.213	37.97	9
ACR-150	2284.40662336857	60.464	37.95	9
INITIAL-1	2348.49847005759	59.952	37.89	9
ACR-173	2267.79743612355	59.932	37.95	9
POC-50	2329.21094044675	60.098	37.9	9

Data and notes for processing E5 samples TP2, Run 9 samples (2), run 10 samples (12), and run 11 samples (2) 20210715

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210715	2230.460292	2224.47	0.269290768	180	CRM opened 20210526

Total Alkalinity Output

SampleID	TA	Mass	Salinity	Run
JUNK1	2270.28371447114	60.173	35	NA
POC-386	2324.0085661087	60.373	38.06	9
POR-354	2309.65549162679	59.868	38.06	9
INITIAL-2	2336.08593365965	59.517	38.15	10
INITIAL-1	2334.68461218021	60.175	38.14	10
POC-44	2316.96398025246	59.912	38.17	10
BLANK-10	2336.38727847286	60.094	38.18	10
POR-79	2257.74172138601	60.193	38.17	10
POC-45	2319.05071812265	60.091	38.18	10
POR-69	2164.33223065006	59.976	38.14	10
POC-68	2321.23543991343	59.537	38.16	10
POR-71	2184.29178281424	60.46	38.17	10
ACR-139	2297.47202192576	59.631	38.17	10
POR-77	2231.06018551181	59.984	38.17	10
ACR-145	2313.15975234594	59.812	38.17	10
POC-55	2314.56794666255	60.002	38.17	11
POR-80	2243.56993167095	59.773	38.17	11

Data and notes for processing E5 samples TP2, Run 11 samples (8), run 20210811

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20210811	2221.057299	2224.47	-0.153416354	180	CRM opened 20210720

Total Alkalinity Output

SampleID	TA	Mass	Salinity
JUNK1	1934.9754608898	60.346	35
BLANK-11	2324.15793070253	59.752	39.4
POC-56	2306.07052131465	59.896	39.42
POC-40	2296.8754717866	60.336	39.42
INITIAL-1	2336.55412418495	59.76	39.41
ACR-178	2269.5758792525	59.807	39.42
POR-78	2159.98302587912	60.382	39.41
POR-70	2181.20006133922	59.548	39.42
ACR-190	2284.78864861159	59.863	39.44

Data and notes for processing E5 samples TP2, Run 11 samples (1), Run 12 samples (7) 20211110

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20211110	2220.494702	2224.47	-0.17870765	180	CRM opened 20210720

Total Alkalinity Output

SampleID	TA	Mass	Salinity
JUNK1	1844.40951345787	60.018	35
POR-81	2279.69144608703	59.848	39.33
POR-74	2287.49989551604	60.48	39.42
Initial-1	2328.69668652401	60.174	39.59
POC-41	2298.07620701969	60.024	39.44
POC-47	2307.75218797885	60.395	39.51
BK-12	2328.61428528263	60.067	39.43
POR-76	2155.56493361319	60.179	39.53
ACR-140	2277.19768756345	59.914	39.75

Data and notes for processing E5 samples TP2, Run 12 samples (5), Run 13 samples (8) 20220210

Notes

• All samples looked good. Updated timeseries data to urol-e5 GitHub.

Data

CRM Accuracy Data

Date	CRM value	Batch value	% off	Batch #	Notes
20220210	2228.61	2224.47	0.186111748	180	CRM OPENED 20220118

Total Alkalinity Output

SampleID	TA	Mass	Salinity	Run
POR_82	2274.19295228573	60.378	40.55	12
POC_57	2319.26422775354	59.96	40.76	12
INITIAL2	2340.15165922546	60.448	40.73	12
POC_43	2318.78430225776	60.445	40.78	12
ACR_51	2280.9306575261	60.203	40.76	12
INITIAL2.1	2330.66583042499	60.354	40.55	13
INITIAL1	2335.34615075794	59.857	40.69	13
BK_13	2334.90676472687	59.544	40.72	13
POC-53	2318.41849540092	60.117	40.65	13
POR-83	2239.15805644992	59.85	40.61	13
POR-72	2310.663725872	59.733	40.65	13
POR-75	2227.34385113541	60.328	40.63	13
POC-52	2171.03862117238	60.093	40.65	13