Approximately 10-15% of couples who want to conceive suffer from subfertility, while in 30% of these cases, male factors play a role. The level of specific microRNAs in seminal plasma, including those involved in spermatogenesis, can serve as a parameter indicative of subfertility. We first optimized protocols to obtain microRNAs from seminal plasma.

Next, using a test strategy is validated in a group of men, we aim to identify microRNAs whose levels associated with semen motility and concentration. By qPCR, 742 microRNAs are profiled in normozoospermic three samples, three samples of sperm and low sperm motility (asthenozoospermia), and three with semen low concentration (oligozoospermia). MicroRNAs show a significant difference between the groups further validated in a second group consisting of 40 samples with normozoospermia (control group), 47 samples with asthenozoospermia, and 19 samples with oligozoospermia (which is 74% too low motility).

MicroRNA highest results obtained with biofluids RNA extraction kits, with the inclusion of MS2 RNA and proteinase K treatment operator for the protocol, and when 50 uL seminal plasma is used as input. Exosome RNA isolation before extraction does not lead to improved results. In the test cohort, 236 microRNAs can be detected, of which 54 microRNAs showed differences between groups. Five microRNAs analyzed in the validation group. the level of MiR-34b-5p in the control group was significantly higher compared with asthenozoospermia group (p <0.05) and compared with oligozoospermia group (p <0.001).

We optimized the acquisition of seminal plasma microRNA and microRNA levels were identified in relation to sperm concentration and motility. As the latest human and rat studies showed that miR-34 family is a marker of low concentration of semen and is very important in spermatogenesis, seminal plasma miR-34b-5p may be a suitable candidate for further study as a marker of male subfertility.

When Less Is More: Special Capture and Analysis in the Plasma Tumor Exosom Liquid Biopsy Improves Sensitivity for Detection of Multiple androgen receptor Comprehensive Phenotypes in Advanced Prostate Cancer Patients

We evaluate the advantages and reliability of new protocols for enriching tumor extracellular vesicles (EV), allowing blood-based test for noninvasive parallel profiles of some of the androgen receptor (AR) gene changes. Three AR clinically relevant variants associated with response / resistance to standard-of-care treatment (AR-V7 transcripts, a point mutation AR T878A and AR gene amplification) were evaluated by digital PCR in 15 samples from patients affected by Castration-Resistant Prostate Cancer (CRPC).

Plasma is processed to obtain circulating RNA and DNA using protocols based enrichment EVS tumor through immuno-affinity and peptide-affinity compared with the generic extraction kit. Our results indicate that the immuno-affinity enrichment before RNA extraction clearly exceed the generic isolation method in the detection of the AR-V7, also allow for the differences between the respondents (R) and non-responders (NR) patients.The T878A mutation is detected, as a whole, in nine out of 15 samples, and no one approach alone is capable of expressing the mutation in all samples store, indicating that the method employed complementary.

Exosome precipitation from Plasma + Thrombin
EPStep-T	ImmunoStep	5 ml	EUR 403.2

ExoQuick Exosome RNA Column Purification kit
EQ808A-1	SBI	20 preps	EUR 482.4

SeraMir Exosome RNA Purification kit (5ml ExoQuick and 20 exoRNA columns)
RA806A-1	SBI	20 preps	EUR 664.8

Exosome precipitation from Plasma and Serum
EPStep-PS	ImmunoStep	5 ml	EUR 356.4

Thrombin Plasma prep for Exosome precipitation (500 ul at 500U/mL)
TMEXO-1	SBI	100 reactions	EUR 699.6

ExoQuick-LP for lipoprotein pre-clear & exosome isolation
EXOLP5A-1	SBI	5 reactions	EUR 639.6

TumorExoRNA? Tumor-derived exosome immunocapture and RNA extraction kit (20 reactions)
K1221-20	Biovision	each	EUR 1462.8

ExoPure? 0.4 micron Immunobeads (Overall Exosome Isolation, cell media, 20 reactions)
M1030-20	Biovision	each	EUR 1546.8

ExoPure? 1.0 micron Immunobeads (Overall Exosome Isolation, cell media, 20 reactions)
M1031-20	Biovision	each	EUR 1546.8

TumorExoPure? 0.4 micron Immunobeads (Tumor-derived Exosome Isolation, biofluids, 20 reactions)
M1032-20	Biovision	each	EUR 1618.8

TumorExoPure? 1.0 micron Immunobeads (Tumor-derived Exosome Isolation, biofluids, 20 reactions)
M1033-20	Biovision	each	EUR 1618.8

MouseExoPure? 0.4 micron Immunobeads (Overall Exosome Isolation, mouse biofluids, 20 reactions)
M1034-20	Biovision	each	EUR 1495.2

MouseExoPure? 1.0 micron Immunobeads (Overall Exosome Isolation, mouse biofluids, 20 reactions)
M1035-20	Biovision	each	EUR 1495.2

Formalin Solution (20%)
FRL125	ScyTek Laboratories	125 ml	EUR 75.6

Formalin Solution (20%)
FRL500	ScyTek Laboratories	500 ml	EUR 98.4

Formalin Solution (20%)
FRL999	ScyTek Laboratories	1000 ml	EUR 121.2

20% SDS Solution
S0793-050	GenDepot	500ml	EUR 144

20% SDS Solution
S0793-100	GenDepot	2X500ml	EUR 199.2

ExoQuick PLUS Exosome Purification Kit - for serum and plasma
EQPL10A-1	SBI	10 reactions	EUR 694.8

MouseExoPure? 0.4 micron Immunobeads (Overall Exosome Isolation, mouse cell media, 20 reactions)
M1036-20	Biovision	each	EUR 1546.8

MouseExoPure? 1.0 micron Immunobeads (Overall Exosome Isolation, mouse cell media, 20 reactions)
M1037-20	Biovision	each	EUR 1546.8

ExoPure? 1.0 micron Immunobeads (Overall Exosome Isolation, plasma, urine, serum, 20 reactions)
M1039-20	Biovision	each	EUR 1546.8

Proteinase K, Recombinant, 20 mg/ml Solution, Molecular Grade
9211-100	Biovision	each	EUR 1495.2

Proteinase K, Recombinant, 20 mg/ml Solution, Molecular Grade
9211-25	Biovision	each	EUR 510

Proteinase K, Recombinant, 20 mg/ml Solution, Molecular Grade
9211-5	Biovision	each	EUR 196.8

ExoDNAPS? Circulating and Exosome-associated DNA Extraction Kit (Human Plasma/Serum, 20 reactions)
K1230-20	Biovision	each	EUR 861.6

ExoDNAUC? Circulating and Exosome-associated DNA Extraction Kit (Urine/Cell Media, 20 reactions)
K1231-20	Biovision	each	EUR 861.6

ExoQuick-TC PLUS Exosome Purification Kit - for tissue culture media
EQPL10TC-1	SBI	10 reactions	EUR 694.8

HSA Solution 20% (cGMP)
AK8214-0100	Akron Biotech	100mL	Ask for price

Arabinose induction solution (20%)
MN850A-1	SBI	20 ml	EUR 152.4

Glycerol, 20% Sterile Solution
GR103065	Genorise Scientific	100 mL	EUR 70.8

100ML SDS Solution 20%
NAT1228	Scientific Laboratory Supplies	100ML	EUR 36.82

450ML SDS Solution 20%
NAT1230	Scientific Laboratory Supplies	450ML	EUR 90.06

SDS, 20% Liquid Solution
18-234	Genesee Scientific	500ml/Unit	EUR 334

SDS, 20% Liquid Solution
18-234B	Genesee Scientific	1000ml/Unit	EUR 390

Puromycin DiHCl Solution (10 mg/mL in 20 mM HEPES)
P025-P026-P025-5x1ML	TOKU-E	P025-5 x 1 mL	EUR 148.8

Puromycin DiHCl Solution (10 mg/mL in 20 mM HEPES)
P026-P026-20ML	TOKU-E	P026-20 mL	EUR 337.2

TotalExoRNA? Overall exosome immunocapture and RNA extraction kit (Biological Fluids/Cell Media, 20 reactions)
K1220-20	Biovision	each	EUR 1462.8

Dispase Solution, 1mg/ml
CA092-010	GenDepot	100ml	EUR 134.4

Transferrin Solution, 10mg/ml
CA205-005	GenDepot	5x1ml	EUR 138

cleaning solution 500 ml
B530	Consort	ea	EUR 36

Kanamycin solution (50mg/ml)
MN800A-1	SBI	20 ml	EUR 152.4

Gentamicin Solution, Gentamicin Solution, 50 mg/mL
CCM1123-010	Bio Basic	10 mL	EUR 76.86

TBE 10X Liquid Solution Ultra Pure Grade Solution
20-196	Genesee Scientific	4 Liters/Unit	EUR 400

ExoQuick Exosome Isolation and RNA Purification kit (for Serum and Plasma)
EQ806A-1	SBI	20 preps	EUR 664.8

ExoQuick Exosome Isolation and RNA Purification kit (for Tissue Culture Media)
EQ806TC-1	SBI	10 preps	EUR 664.8

ExoQuick TC 10ml
EXOTC10A-1	SBI	10 ml	EUR 478.8

ExoQuick TC 50ml
EXOTC50A-1	SBI	50 ml	EUR 1466.4

99445-20 DCT 20 X 150MM
99445-20	CORNING	250/pk	EUR 142.8
Description: Disposable Culture Tubes; DCT's, CGW

20 x Synovial Fluid Solution
GR103059	Genorise Scientific	100 mL	EUR 84

SDS, 20%(w/v) solution
SD8119	Bio Basic	100ml	EUR 76.7

Albumin, Bovine 20mg/ml solution
CH006	ABM	1.0 ml	EUR 170.4

Albumin, Bovine 20mg/ml solution
CH007	ABM	5.0 ml	EUR 254.4

Chloramphenicol Solution (50 mg/ml)
C090-10ML	TOKU-E	10 mL	EUR 70.8

Chloramphenicol Solution (50 mg/ml)
C090-10x10ML	TOKU-E	10 x 10 mL	EUR 349.2

Sodium Selenite Solution,100mg/ml
CA056-010	GenDepot	100ml	EUR 121.2

DNase I Solution, 1mg/ml
CA095-005	GenDepot	5x1ml	EUR 165.6

BSA Blocking Solution, 2 mL
BLK-002	Cusabio	2 mL	EUR 106.21
Description: The BSA solution is blocking gold nanoparticles following conjugation reactions.

BSA Blocking Solution, 10 mL
BLK-010	BioAssayWorks	10 mL	EUR 202.3
Description: The BSA solution is blocking gold nanoparticles following conjugation reactions.

pepsine cleaning solution 250 ml
B231	Consort	ea	EUR 54

pepsine cleaning solution 1000 ml
B232	Consort	ea	EUR 118.8

RNase A Solution (10mg/ml)
RB0474	Bio Basic	1ml	EUR 66.26

ExoDNAPS? Conc Exosome DNA Extraction Kit
K1243-20	Biovision	each	EUR 992.4

ExoDNAUC? Conc Exosome DNA Extraction Kit
K1244-20	Biovision	each	EUR 992.4

LiqBiopsy Tumor Exosome DNA Kit I
K1255-20	Biovision	each	EUR 1201.2

LiqBiopsy Tumor Exosome DNA Kit II
K1256-20	Biovision	each	EUR 1266

DNA Precipitation Buffer
D106	Cygnus Technologies	55 ml	EUR 210
Description: DNA Precipitation Buffer by Cygnus Technologies is available in Europe via Gentaur.

TMB for Precipitation
42R-TB100x	Fitzgerald	1000 ml	EUR 645.6
Description: High sensitivity TMB Substrate for Precipitation assays

EpiNext Hi-Fi cDNA Synthesis Kit
P-9004	EpiGentek	EUR 381.90 EUR 227.70	20 Samples 20 Reactions

20(S)-NotoginsenosideR2
N2028-20	ApexBio	20 mg	EUR 756
Description: Extracted from Pseudo-ginseng;Store the product in sealed, cool and dry condition

ExoPure? Reagent (Overall Exosome Isolation, biological fluids)
M1001-20	Biovision	each	EUR 1260

Human Serum Albumin protein (20% Solution)
30-1138	Fitzgerald	50 mL	EUR 632.4
Description: Human Serum Albumin, 20% solution

Potassium Iodide 20% Solution w/v
KI20WV1	Scientific Laboratory Supplies	1L	EUR 72.96

Potassium Thiocyanate Solution 20% w/v
KT20WV1	Scientific Laboratory Supplies	1L	EUR 74.1

Barium Chloride Solution 20% w/v
WTR081	Scientific Laboratory Supplies	1L	EUR 67.26

50 ML KANAMYCIN SULFATE, LIQUID, 5,000 µG/ML SOLUTION
30-006-CF	CORNING	50 mL/pk	EUR 117.6
Description: Media Catalog; Cell Culture Reagents

Ethidium Bromide, 10mg/ml Stock Solution
E0205-001	GenDepot	10ml	EUR 105.6

Ethidium Bromide, 10mg/ml Stock Solution
E0205-002	GenDepot	20ml	EUR 153.6

Fibronectin, Bovine, Stabilized Solution , 1mg/ml
CA082-001	GenDepot	1ml	EUR 375.6

Fibronectin, Human, Stabilized Solution, 1mg/ml
CA083-001	GenDepot	1ml	EUR 392.4

Vitronectin , Human , Stabilized Solution, 0.5mg/ml
CA085-050	GenDepot	50ul	EUR 250.8

Papain Solution In PBS, 1mg/ml
CA093-002	GenDepot	20ml	Ask for price

Insulin Solution, 1mg/ml , Animal Free
CA204-005	GenDepot	5x1ml	EUR 138

500 ML L-GLUTAMINE 200MM SOLUTION
25-005-CV	CORNING	500 mL/pk	EUR 116.4
Description: Media Catalog; Cell Culture Reagents

100 ML SODIUM BICARBONATE, 7.5% SOLUTION
25-035-CI	CORNING	100 mL/pk	EUR 69.6
Description: Media Catalog; Cell Culture Reagents

buffer solution 500 ml 4.00 ph
B504	Consort	ea	EUR 36

buffer solution 500 ml 7.00 ph
B507	Consort	ea	EUR 36

buffer solution 500 ml 10.01 ph
B510	Consort	ea	EUR 36

buffer solution 500 ml 12.00 ph
B512	Consort	ea	EUR 36

calibration solution 500 ml 200ms/cm
B566B	Consort	ea	EUR 54

buffer solution 50 ml 4.00 ph
B004	Consort	ea	EUR 25.2

buffer solution 50 ml 7.00 ph
B007	Consort	ea	EUR 25.2

buffer solution 50 ml 10.00 ph
B010	Consort	ea	EUR 25.2

calibration solution 50 ml 200ms/cm
B066	Consort	ea	EUR 28.8

Linear Poly Acrylamide Solution (5mg/ml)
LK548	Bio Basic	1ml	EUR 75.66

Hemocyanin-Keyhole Limpet (KLH) subunits, solution
6287-20	Biovision	each	EUR 418.8

20(R)Ginsenoside?Rh1
N2192-20	ApexBio	20 mg	EUR 616.8
Description: Extracted from Panax ginseng C. A. Mey. dried roots;Store the product in sealed, cool and dry condition

20(R)Ginsenoside Rg2
N2195-20	ApexBio	20 mg	EUR 268.8
Description: Extracted from Panax ginseng C. A. Mey. dried roots;Store the product in sealed, cool and dry condition

20(R)Ginsenoside Rg3
N2198-20	ApexBio	20 mg	EUR 616.8
Description: Extracted from Panax ginseng C.A.Mey. roots;Store the product in sealed, cool and dry condition

FGF-20, human recombinant
4589-20	Biovision	each	EUR 332.4

D(+)Glucose 20 % 1000 ml
12-9165-5	Medicago	1000 ml	EUR 170.4

RNaseA50 mg/ml -20 C
FYG106-130UL	Yeastern Biotech	130 µl	Ask for price

RNaseA50 mg/ml -20 C
FYG106-200UL	Yeastern Biotech	200 µl	Ask for price

RNaseA50 mg/ml -20 C
FYG106-50UL	Yeastern Biotech	50 µl	Ask for price

Dubos Med Albu 20 Ml
215334	Scientific Laboratory Supplies	PK10	EUR 134.52

SeraMir Exosome RNA Purification kit (10ml ExoQuick-TC and 10 exoRNA columns) 10 preps
RA806TC-1	SBI	10 preps	EUR 664.8

Reagecon coloured buffer solution - pH at 20 C - pH 4.00 +/- 0.01 - 500 mL bottle
DD440460	Scientific Laboratory Supplies	EACH	EUR 33.06

Reagecon coloured buffer solution - pH at 20 C - pH 10.00 +/- 0.01 - 500 mL bottle
DD440464	Scientific Laboratory Supplies	EACH	EUR 33.06

EtBr Destaining Bag Kit Activation Solution Included
20-277	Genesee Scientific	25 Bags/Unit	EUR 394

Chicken IgY precipitation reagent
85R-1010	Fitzgerald	4 ml	EUR 664.8
Description: Chicken IgY precipitation reagent for use in immunoassays

PEG Virus Precipitation Kit
K2190-200	ApexBio	200 preparations	EUR 1270.8

PEG Virus Precipitation Kit
K2190-50	ApexBio	50 preparations	EUR 486

50ML Protein Precipitation Kit
NAT1254	Scientific Laboratory Supplies	50ML	EUR 237.12

PEG Virus Precipitation Kit
K904-200	Biovision	each	EUR 1201.2

PEG Virus Precipitation Kit
K904-50	Biovision	each	EUR 451.2

Pluronic f-127 20% solution in DMSO
59004	Biotium	1ML	EUR 79.2
Description: Minimum order quantity: 1 unit of 1ML

Hoechst 33258 *20 mM solution in water*
17525	AAT Bioquest	5 mL	EUR 158.4

Hoechst 33342 *20 mM solution in water*
17535	AAT Bioquest	5 mL	EUR 158.4

Hoechst 34580 *20 mM solution in water*
17538	AAT Bioquest	100 uL	EUR 262.8

Sodium dodecyl sulfate, 20% solution in water
GD0990-100ML	Glentham Life Sciences	100 ml	EUR 64.8

Sodium dodecyl sulfate, 20% solution in water
GD0990-1L	Glentham Life Sciences	1 l	EUR 151.2

Sodium dodecyl sulfate, 20% solution in water
GD0990-250ML	Glentham Life Sciences	250 ml	EUR 84

Sodium dodecyl sulfate, 20% solution in water
GD0990-2L	Glentham Life Sciences	2 l	EUR 246

Tween® 20, MegaPure? Detergent, 10% Solution
2125-100	Biovision	each	EUR 242.4

Tween® 20, MegaPure? Detergent, 10% Solution
2125-50	Biovision	each	EUR 176.4

Tween® 20, MegaPure? Detergent, 10% Solution
2125-500	Biovision	each	EUR 836.4

Tween® 20, MegaPure? Detergent, 10% Solution
2125-set	Biovision	each	EUR 222

20 x ELISA Wash Buffer
GR103014-20	Genorise Scientific	1 L	EUR 189.6

Hanks Balanced Salt Solution 500ml
20-021-CVR	Scientific Laboratory Supplies	PK6	EUR 221.16

ExoPure? 0.4 micron Immunobeads (Overall Exosome Isolation, plasma, urine, serum)
M1038-20	Biovision	each	EUR 1546.8

Puromycin Solution, 10mg/ml , Cell Cultere Tested
CR026-001	GenDepot	10ml	EUR 241.2

Fetuin Solution, 1mg/ml in DMEM-F12
CA065-005	GenDepot	5x1ml	EUR 116.4

CometAssay Lysis Solution (2 x 500 mL)
4250-050-01	Biotechne	1 L	EUR 219.6

PBS solution pH 7.4, 1000 ml/bottle
10-9402-10	Medicago	1000 ml/bottle	EUR 193.2

dsGreen Gel Staining Solution, 10000×, 0.5 mL
10010	Lumiprobe	0.5 mL	EUR 106.8

calibration solution 500 ml 0.01 m kcl
B560	Consort	ea	EUR 36

calibration solution 500 ml 0.1 m kcl
B561	Consort	ea	EUR 36

calibration solution 500 ml 1 m kcl
B562	Consort	ea	EUR 36

calibration solution 50 ml 0.01 m kcl
B060	Consort	ea	EUR 25.2

calibration solution 50 ml 0.1 m kcl
B061	Consort	ea	EUR 25.2

calibration solution 50 ml 1 m kcl
B062	Consort	ea	EUR 25.2

redox standard solution 50 ml 124 mv
B071	Consort	ea	EUR 26.4

redox standard solution 50 ml 358 mv
B072	Consort	ea	EUR 26.4

redox standard solution 500 ml 124 mv
B571	Consort	ea	EUR 43.2

redox standard solution 500 ml 358 mv
B572	Consort	ea	EUR 43.2

Spectrophotometric Standard Protein Solution, 3 mg/ml
GR103032	Genorise Scientific	5 x 1 mL	EUR 67.2

X-Gal Solution, 40 mg/ml concentrate
X0321-005	GenDepot	5X1ml	EUR 138

X-Gal Solution, 40 mg/ml concentrate
X0321-020	GenDepot	20ml	EUR 162

TAE 50X Liquid Concentrate Ultra Pure Grade Solution
20-193	Genesee Scientific	1.6 Liters/Unit	EUR 405

TAE 50X Liquid Concentrate Ultra Pure Grade Solution
20-194	Genesee Scientific	4 Liters/Unit	EUR 566

TAE 25X Liquid Concentrate Ultra Pure Grade Solution
20-195	Genesee Scientific	1.6 Liters/Unit	EUR 379

Tris-Glycine 10X Liquid Solution Ultra Pure Grade
20-246	Genesee Scientific	4 Liters/Unit	EUR 402

TG-SDS 10X Liquid Solution Ultra Pure Grade
20-247	Genesee Scientific	4 Liters/Unit	EUR 403

Complete Freund's Adjuvant, 20 mg/ml
7024	Chondrex	20 mg/ml x 5 ml	EUR 472.38
Description: Complete Freund's Adjuvant

20UL FILTER TIPS FOR P-20
TF-20	CORNING	1000/pk	EUR 501.6
Description: Filter Tip; Filter Tips Pipette - Axygen

Anti-Cat Whole Serum | RWS-20
RWS-20	Immunology Consultants Laboratory	1.0 mL	EUR 153
Description: Anti-Cat Whole Serum | RWS-20 | Immunology Consultants Laboratory Host: Rabbit Format: Whole Serum Product Type: Primary Antibody Antibody Clonality: Polyclonal

AR amplification CRPC were detected in most samples were analyzed using either a cell-free DNA (cfDNA) or exosome isolation kit (80%). We show that the selective isolation of exosomes enriched subset for the origin of the tumor, rather than the total analysis of exosomes plasma or plasma total nucleic acids, increased sensitivity and specificity to detect specific changes in circulation.