iNtRON Biotechnology DR

Life Science (Reagent)

All products
Extraction Reagent
- DNA
- RNA
- AutoPrep
Amplification Reagent
- PCR
- RT/RT-PCR
- Real-time PCR
- Others
Research Products
Instruments & Others

RNA

Life Science (Reagent) Extraction Reagent RNA

Patho Gene-spin™ DNA/RNA Extraction Kit

Cat.No	Capacity	Inquire
17154	50 col.
17154.2	200 col

Inquire List

Description
TroubleShooting Guide
Related products
Protocol
MSDS

PRODUCT INFORMATION

Description

A product which extracts DNA/RNA easily and quickly from various pathogenic samples infected by virus or bacteria such as plasma, blood, serum, cell-free body fluids, cell and tissue etc.

• Used low concentration of chaotropic salt and better lysis efficiency

• Higher efficiency of lysis without additional additives

• Stable extraction of DNA/RNA from various pathogenic samples

• Be used safely with non-phenol method and no need for EtOH precipitation

• Suitable for sample extraction such as forensic medicine and diagnosis of diseases etc.

The Patho Gene-spin™ DNA/RNA Extraction Kit uses a low concentration of chaotropic salt and has a better lysis efficiency unlike conventional Total RNA Extraction products. Total DNA/RNA can be extracted from various pathogen samples like virus and bacteria, even anticoagulated plasma/blood, serum, cell-free body fluids and pathogenic cell/tissue etc. And it is easy to induce lysis without any additive like mercaptoethanol. In addition, DNA/RNA extraction is faster and more efficient by using columns that silica-gel membrane technology applied. The DNA/RNA isolated by the Patho Gene-spin™ Kit can be used for clinical diagnosis of people and animals infected with diseases caused by viruses and bacteria.

Applications

01Pathogen detection
02PCR or RT-PCR
03Quantitative PCR (qPCR, qRT-PCR)
04cDNA syntehesis
05Infectious disease research etc.

Kit Contents

Contents	Description	Volumes
Lysis Buffer¹		35ml
Binding Buffer		35ml
Washing Buffer A		30ml
Washing Buffer B²	Add 40 ml of EtOH before use	10ml
Elution Buffer		20ml
Spin Columns	Inserted into a collection tubes. (2.0 ml tubes)	50 columns
Instruction Manual		1 sheet

1. Lysis Buffer is composed high concentration of chaotropic salt. Carefully handle it.
2. Washing Buffer B is supplied as concentrates. Add 40 ml of ethanol (96~100 %) according to the bottle label before use.

Technical Data

Examples of amplification after extractions from various pathogenic samples

After diluting 1/10 of various pathogenic samples, total DNA / RNA was extracted with Patho Gene-spin™ DNA/RNA Extraction Kit. Sensitivity activity and amplification rates were checked using Maxime™ PCR PreMix Kit (i-StarTaq) or Maxime™ RT-PCR PreMix Kit.

Observation of amplification efficiency on various PCR/RT-PCR conditions

Total RNA was extracted from pathogenic samples using the Patho Gene-spin™ DNA/RNA Extraction Kit and competitors DNA/RNA extraction products. PCR / RT-PCR amplification with extracted samples shows that the Patho Gene-spin™ DNA/RNA Extraction Kit is suitable for stable amplification.

Citation List

1	International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 5 Number 4 (2016) pp. 659-666	Isolation and Molecular Characterization of Cry Gene for Bacillus thuringiensis Isolated from Soil of Gaza Strip	Azme Dagga1, Mohamed Abdel Aziz2, Abed Al'raoof Al Amnama3, Mervat Al-Sharif4 and Mahmoud El Hindi5*
2	Virology, April 2017Volume 89, Pages 10–13	Isolation of hepatitis E virus genotype 4 from patients with acute cryptogenic hepatitis	Sook-Hyang Jeong, Byung-Joo Park, Yong-Hyun Ki, Yun Suk Choi, Hee-Seop Ahn, Sang-Hoon Ha, In-Soo Choi
3	Plant Disease / September 2016, Volume 100, Number 9 Page 1958	First Report of Papaya leaf curl virus in Papayas in Korea and Recovery of its Symptoms	H.-S. Byun, E.-J. Kil
4	Journal of Entomology and Zoology Studies 2018; 6(1): 529-540	Isolation and sequencing of field isolates of Avian infectious bronchitis virus in Iraq	Ruya Nabih Mohammed Atta and Dr. Aida Bara Allawe
5	European Journal of Plant Pathology, June 2015, Volume 142, Issue 2, pp 419–426	Identification of natural weed hosts of Tomato chlorosis virus in Korea by RT-PCR with root tissues	Eui-Joon Kil, Ye-Ji Lee, Seungchan Cho, Chung-Kyoon Auh, Donggiun Kim, Kyeong-Yeoll Lee, Mi-Kyeong Kim, Hong-Soo Choi, Chang-Seok Kim Email author , Sukchan Lee
6	Scientific Reports volume 6, Article number: 19013 (2016) doi:10.1038/srep19013	Tomato yellow leaf curl virus (TYLCV-IL): a seed-transmissible geminivirus in tomatoes	Eui-Joon Kil, Sunhoo Kim, Ye-Ji Lee, Hee-Seong Byun, Jungho Park, Haneul Seo, Chang-Seok Kim, Jae-Kyoung Shim, Jung-Hwan Lee, Ji-Kwang Kim, Kyeong-Yeoll Lee, Hong-Soo Choi & Sukchan Lee
7	Journal of Virological Methods Volume 232, June 2016, Pages 12-15	A high-resolution melting (HRM) assay for the differentiation between Israeli field and Neethling vaccine lumpy skin disease viruses	Sophia Menasherow 1, Oran Erster 1, Marisol Rubinstein-Giuni, Anita Kovtunenko, Evgeny Eyngor, Boris Gelman, Evgeny Khinich, Yehuda Stram
8	Vet World. 2018 Sep; 11(9): 1262–1265. Published online 2018 Sep 15. doi: 10.14202/vetworld.2018.1262-1265	Occurrence of Toxoplasma gondii in raw goat, sheep, and camel milk in Upper Egypt	Nagah M. Saad,1 Asmaa A. A. Hussein,2 and Rania M. Ewida3
9	Vol 8 No 2 (2018): April	Incidence of Cronobacter sakazakii in Dairy-based Desserts	Nagah M. Saad, Rania Mohamed Ewida Rania, M. Ewida
10	European Review for Medical and Pharmacological Sciences	SARS-CoV-2 RNA contamination on surfaces of a COVID-19 ward in a hospital of Northern Italy: what risk of transmission?	M. D’ACCOLTI1,2, I. SOFFRITTI1,2, A. PASSARO3,4, G. ZULIANI3,4, P. ANTONIOLI5, S. MAZZACANE2, R. MANFREDINI3,6, E. CASELLI1,2
11	Wiley Online Library	Saliva samples for detection of SARS-CoV-2 in mildly symptomatic and asymptomatic patients	Emin Ediz Tutuncu,Didem Ozgur,Murat Karamese,
12	Wiley Online Library	Long-term follow-up of convalescent pigs and their offspring after an outbreak of acute African swine fever in Vietnam	Taehwan Oh,Tien Manh Nguyen,Tram Thi Ngoc Ngo,Danh Thinh,Trang Thi Phuong Nguyen,Luc Duc Do,Duy Tien Do,
13	Avian Disease	Molecular Detection of a Novel Fowl Adenovirus Serotype-4from an Outbreak of Hepatitis Hydropericardium Syndrome	Hesham Sultan; Abd-Elsatar Arafa; Amany Adel; Karim Selim; Mohamed Hossiny; Shaimaa Talaat
14	bioRxiv	Concentration and quantification of Tilapia tilapinevirus from water using a simple iron flocculation coupled with probe-based RT-qPCR	Suwimon Taengphu, Pattanapon Kayansamruaj, Yasuhiko Kawato, Jerome Delamare-Deboutteville, Chadag Vishnumurthy Mohan, Ha Thanh Dong, Saengchan Senapin
15	BIOSCIENCES BIOTECHNOLOGY RESEARCH ASIA, March 2022. Vol. 19(1), p. 215-229	Screening and Optimisation of the Biodegradation Potential for Low Density Polyethylene (LDPE) Films by Fusarium Equiseti and Brevibacillus Parabrevis	Sally A. Ali1 *, Shimaa Zakarya2 and Shimaa Khaled2
16	British Poultry Science Received 09 Jan 2022, Accepted 22 May 2022, Accepted author version posted online: 06 Jul 2022, Published online: 25 Jul 2022	Development of real time reverse transcription loop-mediated isothermal amplification assay for rapid detection of genotype VII of Newcastle disease viruses	K. Selim,A. AdelORCID Icon,S. EidORCID Icon &M. Shahein
17	Wiley Online Library, 12 April 2022 https://doi.org/10.1111/tbed.14551	Emergence of clade 2.3.4.4b novel reassortant H5N1 high pathogenicity avian influenza virus in South Korea during late 2021	Mingeun Sagong,Yu-Na Lee,San Song,Ra Mi Cha,Eun-Kyoung Lee,Yong-Myung Kang,Hyun-Kyu Cho,Hyun-Mi Kang,Youn-Jeong Lee,Kwang-Nyeong Lee
18	Reserch Square July 1st, 2022, DOI: https://doi.org/10.21203/rs.3.rs-1804349/v1	Antiviral activity of canine interferon lambda 3 expressed using a recombinant adenovirus against canine coronavirus, canine parvovirus, and canine distemper virus	Dong-Hwi Kim, Sang-Hoon Han, Da-Yoon Kim, Jae-Hyeong Kim, Joong-Bok Lee, Seung-Yong Park, Chang-Seon Song In-Soo Choi, Sang-Won Lee
19	Research Square	First report of molecular identification of Phytophthora infestans causing potato late blight in Yemen	Amira Ali Al Harethi, Qais Y. Abdullah, Hala J. Al Jobory, Ramadan A. Arafa
20	Veterinary Research Communications (2023)	Emergence of a novel intergenic region (IGR) IV variant of african swine fever virus genotype II in domestic pigs in Vietnam	Nguyen Tuan Anh Mai, Van Phai Dam, Ki-Hyun Cho, Van Tam Nguyen, Nguyen Van Tuyen, Thi Lan Nguyen, Aruna Ambagala, Jee-Yong Park & Van Phan Le
21	Research Square, 2023	Enhancing durability and Sustainable Preservation of Egyptian Stone Monuments Using metabolites produced by Streptomyces exfoliatus	Basma T. Abd-Elhalim, Bahaa A. Hemdan, Salwa M. El-Sayed, Mahgoub A. Ahmed
22	Enfermedades infecciosas y microbiologia clinica (English ed.) Volume 41, Issue 4, April 2023, Pages 235-237	Low risk of environmental contagion by SARS-CoV-2 in non-sanitary spacesBajo riesgo de contagio ambiental por SARS-CoV-2 en espacios no sanitarios	Sonia Ragull a, Alba Núñez-Gómez a, M. Carmen Aretxalde b, Nieves Zabala b, Noemí Párraga-Niño a c, Miquel Sabrià a c
23	Research Article, 2023	Molecular and serological investigation of Lawsonia intracellularis in weanling foals in Türkiye	Kemal METİNER, Alper METE2, Erdal EROL3
24	Scientific Reports volume 13, Article number: 6691 (2023)	Characterization and control of Rhizoctonia solani affecting lucky bamboo (Dracaena sanderiana hort. ex. Mast.) using some bioagents	Taghreed F. M. Abdel-Rahman, Ahmed Abdel-Megeed & Mohamed Z. M. Salem
25	Enfermedades Infecciosas y Microbiología Clínica Volume 41, Issue 4, April 2023, Pages 235-237	Bajo riesgo de contagio ambiental por SARS-CoV-2 en espacios no sanitariosLow risk of environmental contagion by SARS-CoV-2 in non-sanitary spaces	Sonia Ragull a, Alba Núñez-Gómez a, M. Carmen Aretxalde b, Nieves Zabala b, Noemí Párraga-Niño a c, Miquel Sabrià a c

TroubleShooting Guide

QWhat is the yield of nucleic acid extraction from normal serum samples?: AAccording to some literature reports, there are genome-derived substances from 500 to 1000 in serum or plasma samples derived from healthy individuals. In case of free-circulating DNA in plasma, it is known that it presents at the level of 1-100 ng/ml. This is a fairly large range, so it is practically impossible or insignificant to infer an average yield.

QIs it possible to extract viral nucleic acids from cell culture supernatant?: AYes, it can be used to extract viral nucleic acids from cell culture supernatant in case of following the protocol of this product.

QI want to know the yield of a small amount of DNA or RNA. Is there other way except for using A260 absorbance?: AThe Patho Gene-spin™ DNA/RNA Extraction Kit extracts whole nucleic acids present in a sample. This means that the product extracts not only nucleic acids from the pathogen in the sample but also nucleic acids from the host. Therefore, measuring the absorbance like A260 is not the preferred method. In addition, in the case of a small amount of nucleic acids less than 10 ng / μl, the range of error is large, so it is somewhat inaccurate to trust the measured value. In this case, it is preferable to perform Real-time PCR or Real-time RT-PCR or quantify fluorescent dyes binding to nucleic acids. Of course, since the detection of pathogens belongs to a qualitative study prior to quantification, it is more important to carry out fresh storage of the sample, rapid extraction and subsequent analysis, and it is not meaningful to measure the yield of extracted nucleic acids before use.

QNon-specific bands seem to be increased with the sensitivity of extraction. Is there a way to improve it?: AUnlike serum and plasma samples, whole blood, buffy-coat and tissue emulsion contain large amounts of cells. The Patho Gene-spin™ DNA/RNA Extraction Kit maximizes the efficiency of nucleic acid extraction through effective dissolution of the sample. In result, overall yield is improved. In the case of samples with high cell contents, the amount of nucleic acids derived from the host is also large. Therefore, you can get clear amplification results if template usage is reduced to 1/10 level when non-specific amplification is observed during PCR analysis.