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V5009 One-step Probe RT-qPCR Kit V2

Products & Applications > Molecular Diagnostic Raw Materials > V5009 One-step Probe RT-qPCR Kit V2

V5009 One-step Probe RT-qPCR Kit V2

Price： $320.00~4462.00

Cat. No./Sp：

V5009 (200 rxns) V5010 (5000 rxns)

Product details

Product Introduction

The One-step Probe RT-qPCR Kit V2 is designed for quantitative PCR detection using RNA as a template, such as viral RNA. Using gene-specific primers (GSP), reverse transcription and qPCR reactions are performed in a single tube, requiring no additional tube opening/pipetting, significantly increasing detection throughput and reducing the risk of contamination. This Kit introduced dUTP/UDG system. Heat-labile UDG can rapidly degrade the pollutants containing U at room temperature. When reverse transcription is performed at 55°C, heat-labile UDG will be inactivated rapidly without affecting the efficiency and sensitivity of RT-qPCR. Thermostable reverse transcriptase can maintain stable activity at 55°C. The hot-start antibody-modified fast DNA polymerase and optimized buffer system ensure the extremely high sensitivity and specificity of the One-Step Probe RT-qPCR Kit V2. The kit is available as a convenient Master Mix. 2X RT-qPCR Mix contains an optimized buffer system and dNTP/dUTP Mix, which is suitable for high-specificity detection systems with fluorescent labeled probes such as TaqMan®.

Selection Guide of One-Step Kit (probe based):

Cat. No.	V5001/V5002	V5005/V5006	V5005L/V5006L	V5007/V5008	V5007L/V5008L	V5009/V5010	V5011
Product Name	One-step Probe RT-qPCR Kit	DSPath^TM 4X One-Step Multiplex Master Mix	DSPath^TM 4X One-Step Multiplex Master Mix (Lyophilized)	2X One Step Prime RT-qPCR Mix	2X One Step Prime RT-qPCR Mix (Lyophilized)	One-step Probe RT-qPCR Kit V2	One-step Probe RT-qPCR Kit V3
Mixture Format	Enzyme Mix+2X Buffer Mix	4X All-in Mix	4X All-in Lyophilized Powder	2X All-in Mix	2X All-in Lyophilized Powder	Enzyme Mix+2X Buffer Mix	Enzyme Mix+5X Buffer Mix
Mode/Time	S: ~120 min	F: ~60 min S: ~120 min	F: ~60 min S: ~120 min	F: ~60 min S: ~120 min	F: ~60 min S: ~120 min	F: ~60 min S: ~80 min	F: ~45 min S: ~80 min
RT TEMP	50~55°C	48~55°C	48~55°C	48~55°C	48~55°C	50~55°C	50~55°C
PCR Enzyme	Antibody-modified hotstart Taq	Antibody-modified hotstart Taq	Antibody-modified hotstart Taq	Antibody-modified hotstart Taq	Antibody-modified hotstart Taq	Antibody-modified hotstart Taq	Antibody-modified hotstart Taq
Heat-labile UDG	+	+	+	+	+	+	+
Sensitivity	Very high	Very high	Very high	Very high	Very high	Ultra high	Ultra high

S: standard mode; F: fast mode

V5005, V5009, and V5011 are recommended

Application

• Probe gene expression analysis

• Probe Low-copy gene detection

• Probe microarray validation

• Probe gene knockdown validation

Features

• Fast start-up, 30s hot start

• Reverse transcription at 55 °C to accommodate complex RNA templates

• Contains heat-labile UDG to reduce false-positive contamination

• High sensitivity for detection of low-abundance genes

• High stability, unchanged performance at 4°C and 37°C for one week

• The reaction system can be formulated up to 30 minutes in advance without changing performance

• Saves time by supporting fast program

Experimental Cases

A. Sensitivity: The LOD of Novel Coronavirus is up to 200 copies/ml

B. Stability: when stored at 4°C and 37°C for a week, the performance does not change significantly:

C. Performance comparison test with other suppliers:

l compared with Chinese manufacturers: better performance than the supplier N and supplier B.

Test sample: pseudovirus of SARS-CoV-2

l Comparison with international suppliers: no matter the use of standard mode, or fast mode, performance is not lost to international well-known brands.

Test sample: pseudovirus of SARS-CoV-2

l No immediate reaction is required after the reaction system is formulated. The system can be stored at RT before reaction, and the performance has no obvious effect within 30 minutes.

Test sample: pseudovirus of SARS-CoV-2

Sheet 1: perform reaction immediately after the reaction system is ready

Dilution fold		1			2			10
Supplier	Gene	CT	SD	n	CT	SD	n	CT	SD	n
GDSBIO	N	35.53	0.49	10	36.86	0.58	10	39.25	0.75	8
B	N	36.33	0.56	10	37.54	0.89	10	39.69	1.29	5
GDSBIO	RNaseP	35.97	0.51	10	36.41	0.89	10	38.86	0.62	9
B	RNaseP	35.51	0.46	10	36.69	1.17	10	38.34	0.98	9
GDSBIO	ORF1ab	35.22	0.74	10	36.37	1.09	10	37.29	0.63	4
B	ORF1ab	35.74	0.66	10	37.23	2.38	8	38.52	0.20	2

Sheet 2: perform reaction 30min after the reaction system is ready and placed at RT

Dilution fold		1			2			10
Supplier	Gene	CT	SD	n	CT	SD	n	CT	SD	n
GDSBIO	N	34.88	0.56	10	35.48	0.59	10	38.18	0.44	5
B	N	35.65	0.81	10	38.24	2.67	7	39.47	2.64	3
GDSBIO	RNaseP	35.80	0.44	10	35.76	0.59	10	37.88	0.73	8
B	RNaseP	34.80	0.27	10	36.00	1.06	10	39.93	2.17	7
GDSBIO	ORF1ab	35.65	0.62	10	37.53	1.06	8	38.69	0.72	7
B	ORF1ab	null	null	0	null	null	0	null	null	0

The performance of V5009 does not change when the reaction system is prepared 30 minutes in advance. However, the testing performance of Company B product decreased significantly after it was left at room temperature for 30min after reaction system was ready.

Components

Component

V5009

(200 rxns)

V5010

(5000 rxns)

2X One-step U⁺ Mix

760 μl × 4

38.4 ml × 2

One-step U⁺ Enzyme Mix V2

308 μl

7.7 ml