aprofarm

BDNF Val66Met Polymorphism, the Allele-Specific Analysis by QRT-PCR – a Novel Protocol

Molecular Blood Group Screening in Donors from Arabian Countries and Iran Using High-Throughput MALDI-TOF Mass Spectrometry and PCR-SSP

Background and aims: Only little is known about blood groups other than ABO blood groups and Rhesus factors in Arabian countries and Iran. During the last years, increased migration to Central Europe has put a focus on the question how to guarantee blood supply for patients from these countries, particularly because hemoglobinopathies with the need of regular blood support are more frequent in patients from that region. Therefore, blood group allele frequencies should be determined in individuals from Arabian countries and Iran by molecular typing and compared to a German rare donor panel.
Methods: 1,111 samples including 800 individuals from Syria, 147 from Iran, 123 from the Arabian Peninsula, and 41 from Northern African countries were included in a MALDI-TOF MS assay to detect polymorphisms coding for Kk, Fy(a/b), Fy<sub>null</sub>, C<sub>w</sub>, Jk(a/b), Jo(a+/a-), Lu(a/b), Lu(8/14), Ss, Do(a/b), Co(a/b), In(a/b), Js(a/b), Kp(a/b), and variant alleles RHCE*c.697C>G and RHCE *c.733C>G. Yt(a/b), S-s-U-, Vel<sub>null</sub>, Co<sub>null</sub>, and RHCE *c.667G>T were tested by PCR-SSP.
<strong class=”sub-title”>
Results: </strong> Of the Arabian donors, 2% were homozygous for the <i>FY</i> *<i>02.01N</i> allele (Fy<sub>null</sub>), and 15.7% carried the heterozygous mutation. However, 0.8% of the German donors also carried 1 copy of the allele. 3.6% of all and 29.3% of Northern African donors were heterozygous for the <i>RHCE</i> *<i>c.733C>G</i> substitution, 0.4% of the Syrian probands were heterozygous for <i>DO</i> *<i>01/DO</i> *<i>01.-05</i>, a genotype that was lacking in German donors.
The egg drop syndrome (76) PCR kit
PCR-V044-48D 50T
EUR 425.80
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of The egg drop syndrome (76)
The egg drop syndrome (76) PCR kit
PCR-V044-96D 100T
EUR 521.50
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of The egg drop syndrome (76)
PCR Mix
L5051100 2.5 ml
EUR 107.00
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.
amfiSure qGreen Q-PCR Master Mix(2X), No Rox
Q5600-001 1ml
EUR 146.00
amfiSure qGreen Q-PCR Master Mix(2X), No Rox
Q5600-002 2x1ml
EUR 207.00
amfiSure qGreen Q-PCR Master Mix(2X), No Rox
Q5600-005 5x1ml
EUR 269.00
amfiSure qGreen Q-PCR Master Mix(2X), No Rox
Q5600-010 10x1ml
EUR 448.00
amfiSure qGreen Q-PCR Master Mix(2X), No Rox
Q5600-050 50x1ml
EUR 1738.00
amfiSure qProbe Q-PCR Master Mix(2X), No Rox
Q5700-001 1ml
EUR 174.00
amfiSure qProbe Q-PCR Master Mix(2X), No Rox
Q5700-002 2x1ml
EUR 266.00
amfiSure qProbe Q-PCR Master Mix(2X), No Rox
Q5700-005 5x1ml
EUR 469.00
amfiSure qProbe Q-PCR Master Mix(2X), No Rox
Q5700-010 10x1ml
EUR 739.00
amfiSure qProbe Q-PCR Master Mix(2X), No Rox
Q5700-050 50x1ml
EUR 2885.00
2×Taq PCR Master Mix(with dye)
K1034-1 1 ml
EUR 73.00
0.2ML THIN WALL PCR TUBES, NO CAP. CLEAR
PCR-02-NC 1000/pk
EUR 318.00
Description: PCR Plates & Tubes; PCR Tubes - Axygen
Ready? PCR Mix
M1127-1000
EUR 512.00
Ready? PCR Mix
M1127-200
EUR 191.00
Rigor? PCR Mix
M1132-200
EUR 321.00
Breeze? PCR Mix
M1134-200
EUR 267.00
Distant? PCR Mix
M1136-200
EUR 294.00
Advance? PCR Mix
M1139-200
EUR 376.00
Egg Drop Syndrome RT PCR kit
RTq-V026-100D 100T
EUR 628.50
  • Contact us in order to know the reactivity of the kit.
Description: A Real-Time PCR kit for detection of Egg Drop Syndrome .
Egg Drop Syndrome RT PCR kit
RTq-V026-150D 150T
EUR 701.00
  • Contact us in order to know the reactivity of the kit.
Description: A Real-Time PCR kit for detection of Egg Drop Syndrome .
Egg Drop Syndrome RT PCR kit
RTq-V026-50D 50T
EUR 532.80
  • Contact us in order to know the reactivity of the kit.
Description: A Real-Time PCR kit for detection of Egg Drop Syndrome .
HotTaq Probe qPCR Mix (no ROX)
BT11002 250rxn
EUR 109.00
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
HotTaq EvaGreen qPCR Mix (no ROX)
BT11102 250rxn
EUR 123.00
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
HotTaq EvaGreen HRM Mix (no ROX)
BT11202 250rxn
EUR 123.00
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
dNTP Mix (1 ml)
9002-1
EUR 153.00
Individual Reaction Mix 1
G065-1 200 reactions
EUR 167.00
PCR-EZ D-PCR MASTER MIX
BS294 100RXN, 100prep
EUR 93.50
  • Product category: PCR Related/PCR Premix (dPCR)/dPCR Premix + Taq
amfiSure PCR Master Mix
P0311-010 2x50 rxns
EUR 126.00
amfiSure PCR Master Mix
P0311-025 5X50 rxns
EUR 165.00
amfiSure PCR Master Mix
P0311-050 10X50 rxns
EUR 219.00
amfiSure PCR Master Mix
P0311-100 10x1ml
EUR 286.00
amfiSure PCR Master Mix
P0311-125 15X50 rxns
EUR 278.00
amfiSure PCR Master Mix
P0311-200 20x1ml
EUR 469.00
amfiSure PCR Master Mix
P0311-250 50x50 rxns
EUR 752.00
amfiSure PCR Master Mix
P0311-500 100x50 rxns
EUR 1258.00
amfiXpand PCR Master Mix
P0331-010 2X50 rxns
EUR 151.00
amfiXpand PCR Master Mix
P0331-025 5X50 rxns
EUR 273.00
amfiXpand PCR Master Mix
P0331-050 10X50 rxns
EUR 448.00
amfiXpand PCR Master Mix
P0331-250 50x50 rxns
EUR 1873.00
Ready? PCR Mix-Dye
M1128-1000
EUR 512.00
Ready? PCR Mix-Dye
M1128-200
EUR 191.00
Image Ready? PCR Mix
M1129-200
EUR 294.00
Robust Ready? PCR Mix
M1130-1000
EUR 615.00
Robust Ready? PCR Mix
M1130-200
EUR 213.00
Rigor? PCR Mix-Dye
M1133-200
EUR 321.00
Breeze? PCR Mix-Dye
M1135-200
EUR 267.00
Distant? PCR Mix-Dye
M1137-200
EUR 294.00
Image Distant? PCR Mix
M1138-200
EUR 321.00
Advance? PCR Mix-Dye
M1140-200
EUR 376.00
Fire Start? PCR Mix
M1141-200
EUR 376.00
Whole Blood PCR Mix
M1143-200
EUR 321.00
7078D 1 PIPET 1ML STERILE STERILE
7078D-1 50/pk
EUR 180.00
Description: Disposable Pipets; Disposable Pipets, Serological, Misc
Egg Drop Syndrome One-Step PCR kit
Oneq-V026-100D 100T
EUR 747.40
  • Contact us in order to know the reactivity of the kit.
Description: Real Time PCR Kit is a screening assay for a rapid and accurate detection of Egg Drop Syndrome.
Egg Drop Syndrome One-Step PCR kit
Oneq-V026-150D 150T
EUR 838.75
  • Contact us in order to know the reactivity of the kit.
Description: Real Time PCR Kit is a screening assay for a rapid and accurate detection of Egg Drop Syndrome.
Egg Drop Syndrome One-Step PCR kit
Oneq-V026-50D 50T
EUR 628.50
  • Contact us in order to know the reactivity of the kit.
Description: Real Time PCR Kit is a screening assay for a rapid and accurate detection of Egg Drop Syndrome.
SensiFAST Lyo-Ready No-ROX Mix, 2x
BIO-11060 10 x 10ml Ask for price
SensiFAST Lyo-Ready No-ROX Mix, 2x
BIO-11061 5ml Ask for price
SensiFAST Probe No-ROX Mix, 50 mL
BIO-86050 5000 rxn Ask for price
SensiFAST SYBR No-ROX Mix, 50 mL
BIO-98050 5000 rxn Ask for price
SYBR Green qPCR Master Mix (No ROX)
HY-K0523 5 mL (500 rxns )
EUR 257.00
Accuris qMax Probe No Rox qPCR Mix
PR2001-N-1000 1 PC
EUR 467.74
  • To order instruments in 115V / US plug please delete the 'E' off the order code.European 2 pin plugs will be supplied as standard, please request UK if required.
7077B PIPET SHRTY 1ML
7077B-1 250/pk
EUR 231.00
Description: Disposable Pipets; Disposable Pipets, Serological, Misc
7078B PIPET SHRTY 1ML
7078B-1 250/pk
EUR 240.00
Description: Disposable Pipets; Disposable Pipets, Serological, Misc
2x GoldStar Best PCR Master Mix (with Dyes), 20 ul/rxn
W0655-1 NULL
EUR NULL
MyTaq Blood PCR Mix, 2x
BIO-25053/S Sample Ask for price
MyTaq Blood PCR Mix, 2x
BIO-25054 250 Reactions Ask for price
amfiSure Prime PCR Master Mix
P1311-025 5X50 rxns
EUR 161.00
amfiSure Prime PCR Master Mix
P1311-050 10X50 rxns
EUR 219.00
amfiSure Prime PCR Master Mix
P1311-125 15X50 rxns
EUR 273.00
amfiSure Prime PCR Master Mix
P1311-250 50x50 rxns
EUR 739.00
amfiSure Prime PCR Master Mix
P1311-500 100x50 rxns
EUR 1238.00
amfiSure Advanced PCR Master Mix
P2311-025 5X50 rxns
EUR 165.00
amfiSure Advanced PCR Master Mix
P2311-050 10X50 rxns
EUR 223.00
amfiSure Advanced PCR Master Mix
P2311-125 15X50 rxns
EUR 282.00
amfiSure Advanced PCR Master Mix
P2311-250 50x50 rxns
EUR 752.00
amfiSure Advanced PCR Master Mix
P2311-500 100x50 rxns
EUR 1279.00
Robust Ready? PCR Mix-Dye
M1131-1000
EUR 615.00
Robust Ready? PCR Mix-Dye
M1131-200
EUR 213.00
Fire Start? PCR Mix-Dye
M1142-200
EUR 376.00
mRNAExpress Tail PCR mix (5uM)
MR-TAIL-PR 100 ul
EUR 164.00
  • Category: MicroRNA Tools
The egg drop syndrome (76) RT PCR kit
RTq-V044-100D 100T
EUR 628.50
  • Contact us in order to know the reactivity of the kit.
Description: A Real-Time PCR kit for detection of The egg drop syndrome (76) .
The egg drop syndrome (76) RT PCR kit
RTq-V044-150D 150T
EUR 701.00
  • Contact us in order to know the reactivity of the kit.
Description: A Real-Time PCR kit for detection of The egg drop syndrome (76) .
The egg drop syndrome (76) RT PCR kit
RTq-V044-50D 50T
EUR 532.80
  • Contact us in order to know the reactivity of the kit.
Description: A Real-Time PCR kit for detection of The egg drop syndrome (76) .
PRICE DROP
110-012 4 x 1000 µl
EUR 133.00
PRICE DROP
110-012L 5x 4 x 1000 µl
EUR 518.00
PRICE DROP
110-012XL 10x 4 x 1000 µl
EUR 939.00
The egg drop syndrome (76) One-Step PCR kit
Oneq-V044-100D 100T
EUR 747.40
  • Contact us in order to know the reactivity of the kit.
Description: Real Time PCR Kit is a screening assay for a rapid and accurate detection of The egg drop syndrome (76).
The egg drop syndrome (76) One-Step PCR kit
Oneq-V044-150D 150T
EUR 838.75
  • Contact us in order to know the reactivity of the kit.
Description: Real Time PCR Kit is a screening assay for a rapid and accurate detection of The egg drop syndrome (76).
The egg drop syndrome (76) One-Step PCR kit
Oneq-V044-50D 50T
EUR 628.50
  • Contact us in order to know the reactivity of the kit.
Description: Real Time PCR Kit is a screening assay for a rapid and accurate detection of The egg drop syndrome (76).
Taq 5x PCR Mix 12.5 mM
BT10402 250rxn
EUR 109.00
Description: High quality Taq polymerase for different PCR variations and downstream applications.
Taq 2x PCR Mix 100 rxn
BA01501 100rxn
EUR 94.00
Description: High quality Taq polymerase for different PCR variations and downstream applications.
HotTaq 2x PCR Mix 100 rxn
BA01503 100rxn
EUR 120.00
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
RedTaq 2x PCR Mix 100 rxn
BA01507 100rxn
EUR 146.00
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.
amfiSure ONE PCR Master Mix(2X)
P7000-005 5x1 ml
EUR 219.00
amfiSure ONE PCR Master Mix(2X)
P7000-010 10x1 ml
EUR 317.00
amfiSure ONE PCR Master Mix(2X)
P7000-050 50x1 ml
EUR 1211.00
amfiSure ONE PCR Master Mix(2X)
P7000-100 100x1 ml
EUR 2102.00
AXYPREP MAG PCR NORMALIZER KIT- 1 ML - (SAMPLE SIZE)
MAG-PCR-NM-1 1/pk
EUR 73.00
Description: Bioscience Mag Beads; Magnetic PCR Normaliser
AXYPREP MAG PCR CLEAN UP KIT- 1 ML - (SAMPLE SIZE)
MAG-PCR-CL-1 1/pk
EUR 58.00
Description: Bioscience Mag Beads; Magnetic PCR Clean Up
Poliovirus 1 (PV1) PCR kit
PCR-H659-48R 50T
EUR 524.40
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Poliovirus 1 (PV1)
Poliovirus 1 (PV1) PCR kit
PCR-H659-96R 100T
EUR 669.40
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Poliovirus 1 (PV1)
Psittacid herpesvirus 1 PCR kit
PCR-V014-48D 50T
EUR 425.80
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Psittacid herpesvirus 1
Psittacid herpesvirus 1 PCR kit
PCR-V014-96D 100T
EUR 521.50
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Psittacid herpesvirus 1
Equine herpesvirus 1 PCR kit
PCR-V144-48D 50T
EUR 425.80
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Equine herpesvirus 1
Equine herpesvirus 1 PCR kit
PCR-V144-96D 100T
EUR 521.50
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Equine herpesvirus 1
Bovine herpesvirus 1 PCR kit
PCR-V182-48D 50T
EUR 425.80
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Bovine herpesvirus 1
Bovine herpesvirus 1 PCR kit
PCR-V182-96D 100T
EUR 521.50
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Bovine herpesvirus 1
Bluetongue virus 1 PCR kit
PCR-V226-48R 50T
EUR 524.40
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Bluetongue virus 1
Bluetongue virus 1 PCR kit
PCR-V226-96R 100T
EUR 669.40
  • Contact us in order to know the reactivity of the kit.
Description: An conventional PCR kit for detection of Bluetongue virus 1
384 WELL PCR MICROPLATE FOR ROCHE 480 LIGHT CYCLER, WHITE (NO SEALING FILM)
PCR-384-LC480-W-NF 10/pk
EUR 240.00
Description: PCR Plates & Tubes; A354 PCR Plate-Axygen
96 WELL PCR MICROPLATE FOR ROCHE 480 LIGHT CYCLER, CLEAR (NO SEALING FILM)
PCR-96-LC480-C-NF 10/pk
EUR 199.00
Description: PCR Plates & Tubes; A354 PCR Plate-Axygen
96 WELL PCR MICROPLATE FOR ROCHE 480 LIGHT CYCLER, WHITE (NO SEALING FILM)
PCR-96-LC480-W-NF 10/pk
EUR 199.00
Description: PCR Plates & Tubes; A354 PCR Plate-Axygen
One-Step BrightGreen qRT-PCR-No Dye
G471-S 100 rxn (20 ul/rxn)
EUR 158.00
One-Step TaqProbe qRT-PCR-No Dye
G493-PS 100 rxn (20 ul/rxn)
EUR 158.00
Smart 2X PCR pre-mix Pfu (30ul)
9K-002-0038 30uL, 30uL
EUR 581.14
  • Product category: PCR Related/PCR Premix/PCR Premix + Pfu
HotStart PCR Master mix (2X, Green Dye)
BS9291 5X1mL, 5ml
EUR 209.50
  • Product category: PCR Related/PCR Premix (qPCR)/qPCR Premix + Taq
HotStart PCR Master mix (2X, Green Dye)
BS9292 1ml
EUR 96.40
  • Product category: PCR Related/PCR Premix (qPCR)/qPCR Premix + Taq
aprofarm
aprofarm
Whereas the <i>KEL</i> *<i>02.06</i> allele coding for the Js(a) phenotype was missing in Germans; 0.8% of the Syrian donors carried 1 copy of this allele. 1.8% of the Syrian but only 0.3% of the German donors were negative for <i>YT</i> *<i>01</i>. One donor from Northern Africa homo-zygously carried the <i>GYPB</i> *<i>270+5g>t</i> mutation, inducing the S-s-U+<sup>w</sup> phenotype, and in 2 German donors a <i>GYPB</i> *<i>c.161G>A</i> exchange, which induces the Mit+ phenotype, caused a <i>GYPB</i> *<i>03</i> allele dropout in the MALDI assay. The overall failure rate of the Arabian panel was 0.4%.

CRISPR-mediated knock-in in the mouse embryo using long single stranded DNA donors synthesised by biotinylated PCR

Successful gene knock-in by CRISPR-Cas9 in the mouse zygote requires three components; guideRNA, Cas9 protein and a suitable donor template, which usually comprises homology flanked insert sequence. Recently, long single stranded DNA (lssDNA) donors have emerged as a popular choice of DNA donor, outperforming dsDNA templates in terms of knock-in efficiency for gene tagging and generating conditional alleles.

 

The generation of these donors can be achieved through several methods that may introduce errors in the sequence, result in poor yields, and contain dsDNA contamination. We have developed our own cost-effective lssDNA synthesis methodology that results in high purity, sequence verified, low contamination lssDNA donors. We provide a detailed methodology on the design and generation of such donors for gene tagging experiments and generating conditional alleles.

BDNF Val66Met Polymorphism, the Allele-Specific Analysis by qRT-PCR – a Novel Protocol

Background: Alteration in brain-derived neurotrophic factor (BDNF) production is a marker of neuropathological conditions, which has led to the investigation of Val66Met polymorphism occurring in the human BDNF gene (BDNF). Presently, there are no reported methods available for the analysis of Val66Met impact on human BDNF functioning.

Purpose: To develop a qRT-PCR protocol for the allele-specific expression evaluation of the Val66Met polymorphism in BDNF

Methods: Using RNA extracted from muscle samples of 9 healthy volunteers (32.9 ± 10.3 y) at rest and following a maximal effort aerobic capacity exercise test, a protocol was developed for the detection of Val66/Met66 allele-specific BDNF expression in Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) – relative to housekeeping genes – and validated by absolute quantification in Droplet Digital Polymerase Chain Reaction (ddPCR).

 

Results: Differences in the relative values of BDNF mRNA were confirmed by ddPCR analysis. HPRT1 and B2M were the most stable genes expressed in muscle tissue among different metabolic conditions, while GAPDH revealed to be metabolic responsive. Conclusion: Our qRT-PCR protocol successfully determines the allele-specific detection and changes in BDNF expression regarding the Val66Met polymorphism.

Standardization of BCR-ABL1 p210 Monitoring: From Nested to Digital PCR

The introduction of tyrosine kinase inhibitors in 2001 as a targeted anticancer therapy has significantly improved the quality of life and survival of patients with chronic myeloid leukemia. At the same time, with the introduction of tyrosine kinase inhibitors, the need for precise monitoring of the molecular response to therapy has emerged. Starting with a qualitative polymerase chain reaction, followed by the introduction of a quantitative polymerase chain reaction to determine the exact quantity of the transcript of interest-p210 BCR-ABL1, molecular monitoring in patients with chronic myeloid leukemia was internationally standardized.

 

This enabled precise monitoring of the therapeutic response, unification of therapeutic protocols, and comparison of results between different laboratories. This review aims to summarize the steps in the diagnosis and molecular monitoring of p210 BCR-ABL1, as well as to consider the possible future application of a more sophisticated method such as digital polymerase chain reaction.

Droplet Digital PCR Analysis of Liquid Biopsy Samples Unveils the Diagnostic Role of hsa-miR-133a-3p and hsa-miR-375-3p in Oral Cancer

Despite the availability of screening programs, oral cancer deaths are increasing due to the lack of diagnostic biomarkers leading to late diagnosis and a poor prognosis. Therefore, there is an urgent need to discover novel effective biomarkers for this tumor. On these bases, the aim of this study was to validate the diagnostic potential of microRNAs (miRNAs) through the analysis of liquid biopsy samples obtained from ten oral cancer patients and ten healthy controls. The expression of four selected miRNAs was evaluated by using droplet digital PCR (ddPCR) in a pilot cohort of ten oral cancer patients and ten healthy donors. Bioinformatics analyses were performed to assess the functional role of these miRNAs.

 

The expression levels of the predicted down-regulated hsa-miR-133a-3p and hsa-miR-375-3p were significantly reduced in oral cancer patients compared to normal individuals while no significant results were obtained for the up-regulated hsa-miR-503-5p and hsa-miR-196a-5p. ROC analysis confirmed the high sensitivity and specificity of hsa-miR-375-3p and hsa-miR-133a-3p. Therefore, both miRNAs are significantly down-regulated in cancer patients and can be used as biomarkers for the early diagnosis of oral cancer. The analysis of circulating miRNAs in a larger series of patients is mandatory to confirm the results obtained in this pilot study.

Leave a Reply

Your email address will not be published. Required fields are marked *