User Manual FavorPrep Viral Nucleic Acid Extraction
Contents
1. cell cultured supernatant Handing time 20 min Elution Volume 50 ul Important Notes 1 Make sure everything is RNase free when handling this system 2 Buffers provided in this system contain irritants Wear gloves and lab coat when handling these buffers 3 Add 1 ml of VNE Buffer to the tube of lyophilized Carrier RNA mix well by vortexing and transfer the mixture to the VNE Buffer when first open Store the Carrier RNA added VNE Buffer at 4 C Brief Procedure 150 ul of sample Lysis VNE Buffer 10 min Ethanol 96 100 Binding i centrifuge Washing Wash Buffer 1 D Wash Buffer 2 X 2 w centrifuge Elution RNase free Water centrifuge General Protocol Please Read Important Notes Before Starting Following Steps _ Transfer 150 ul of sample serum plasma body fluids or cell cultured supernatant into a microcentrifuge tube not provided If the sample volume is more than 150 ul separate it into multiple tubes Add 570 ul of VNE Buffer Carrier RNA added to the sample mix well by vortexing and incubate for 10 minutes at room temperature Make sure that Carrier RNA has been added to the VNE Buffer when first use Add 570 ul of ethanol 96 100 to the sample mixture mix well by plus vortexing Combine a VNE column with a Collection Tube provided Transfer up to 700 ul of sample mixture ethanol added to the VNE Column cent2. VNE Buffer 30 ml 60 ml 180 ml Wash Buffer 1 concentrated 22 ml 44 ml 132 ml Wash Buffer 2 concentrated 20 ml 20 ml X 2 50ml x 2 RNase free Water 6 ml 12 ml 30 ml VNE Column 50 pcs 100 pcs 300 pcs Collection Tube 100 pcs 200 pcs 600 pcs Elution Tube 50 pcs 100 pcs 300 pcs User Manual 1 1 1 Add 30 ml 60 ml 180 ml of ethanol 96 100 to AD Buffer when first open Add 8 ml 16ml 48ml of ethanol 96 100 to Wash Buffer 1 when first open Add 80 ml 200 ml of ethanol 96 100 to Wash Buffer 2 when first open Specification Sample Source Serum Plasma Cell Culture Supernatants Cell Free Body Fluids Sample Size 200 ul Operation time lt 20 min Important Notes 1 Make sure everything is RNase free when handling this system 2 Buffers provided in this system contain irritants Wear gloves and lab coat when handling these buffers 3 For FAVNK002 add 30 ml of ethanol 96 100 to AD Buffer when first open For FAVNK002 1 add 60 ml of ethanol 96 100 to AD Buffer when first open 4 For FAVNK002 add 8 ml of ethanol 96 100 to Wash Buffer 1 when first open For FAVNK002 1 add 16 ml ethanol 96 100 to Wash Buffer 1 when first open 4 Add 80 ml of ethanol 96 100 to each Wash Buffer 2 when first open Brief Procedure Virial Sample Lysis VNE Buffer centrifuge AD Buffer Binding centrifuge Washing Wash Buffer 1 Wash Buffer2 x 2 centrifuge Elution RNase fre
3. FavorPrep Viral Nucleic Acid Extraction Kit User Manual Cat No FAVNK 001 50 Preps FAVNK 001 1 100 Preps FAVNK 001 2 300 Preps For Research Use Only v 0912 Introduction FavorPrep Viral Nucleic Acid Extraction Mini Kit I is desigened for extraction of Viral DNA or RNA from cell free fluides such as serum plasma body fluid and cell cultured supernatant This method first lyses virus by using a chaotropic salt then binds nucleic acid to silica based membranes After washing with ethanol contained wash buffer con taminants and enzyme inhibitors will be removed completely It takes only 20 min for an entire procedure the purified nucleic acid is ready for RT PCR and PCR Kit Contents Cat No FAVNKOO1 FAVNKOO1 1 FAVNKO01 2 preps 50 preps 100 preps 300 preps VNE Buffer 35 ml 70 ml 200 ml Wash Buffer 1 concentrated 22 ml 44 ml 132 ml Wash Buffer 2 concentrated 20 ml 20 mi x 2 50 ml x 2 RNase free Water 6 ml 12 ml 20 ml Carrier RNA 0 4 mg 0 8 mg 2 2 mg VNE Column 50 pcs 100 pcs 300 pcs Collection Tube 100 pcs 200 pcs 600 pcs Elution Tube 50 pcs 100 pcs 300 pcs User Manual 1 1 1 Add 8 ml 16 ml 48 ml of ethanol 96 100 to Wash Buffer 1 when first open For FAVNK001 FAVNK001 1 add 80 ml of ethanol 96 100 to each Wash Buffer 2 when first open For FAVNK001 2 add 200 ml of ethanol 96 100 to each Wash Buffer 2 when first open Specification Sample 150 ul of plasma serum body fluid and
4. VNE Column for 2 min Important step For effective elution make sure that the RNase free Water is dispensed onto the membrane center and is absorbed completely 11 Centrifuge for 2 min to elute the nucleic acid 12 Store nucleic acid at 70 C FavorPrep Viral Nucleic Acid Extraction Kit Il User Manual Cat No FAVNK 002 50 Preps FAVNK 002 1 100 Preps FAVNK 002 2 300 Preps For Research Use Only v 1304 Introduction The Viral Nucleic Acid Extraction Kit isdesigned for purification of total RNA or DNA from cell free samples such as plasma serum urine cell culture super natant or cell free body fluid The method utilises detergents and a chao tropic salt to lyse virus then the nucleic acid in chaotropic salt is bound to the glass fiver matrix of column After washing off the contaminants the purfied nucleic acid is eluted by RNase free water The detection limit for certain viruses depends on the sensitivity of individual PCR or RT PCR assay This proto col is recommended for parallel purification of viral RNA including HCV HIV and HTLV and viral DNA including HBV and CMV Extracted DNA can be used directly for PCR amplification RNA for RT PCR amplification The entire proce dure can be completed in 20 minutes This kit specially is for low viral load specimen Kit Contents Cat No FAVNK002 FAVNKOO2 1 FAVNK002 2 preps 50 preps 100 preps 300 preps AD Buffer concentrated 4ml 8 ml 24 ml
5. e water centrifuge General Protocol Please Read Important Notes Before Starting Following Steps _ Transfer 200 ul of sample serum plasma body fluids and the supernatant of viral infected cell culture into a microcentrifuge tube not provided lf prepared sample is less than 200 ul adjust sample volume to 200ul with PBS not provided Add 500 ul of VNE Buffer to the sample mix well by vortexing Incubate the sample mixture at room temperature for 10 minutes Add 550 ul of AD Buffer ethanol added to the sample mixture and mix well immediately by plus vortexing Make sure that ethanol has been added into AD Buffer when first open Combine a VNE column with a 2 ml Collection tube Transfer up to 750 ul of sample mixture to the VNE column Cenirifuge at 8 000 x g for 1 minute then discard the flow through Combine the VNE Column with the used Collection Tube Transfer the rest of sample mixture to the VNE Column Cenirifuge at 8 000 x g for 1 min then discard the flow through and the Collection Tube Combine the VNE Column with a new Collection Tube provided Add 500 ul of Wash Buffer 1 ethanol added to the VNE Column centrifuge at 8 000 x g for 1 min then discard the flow through Combine the VNE Column with the used Collection Tube Make sure that ethanol 96 100 has been added into Wash Buffer 1 when first open Add 750 ul of Wash Buffer 2 ethanol added to VNE Col
6. rifuge at 8 000 x g for 1 min then discard the flow through Combine the VNE Column with the used Collection Tube Transfer the rest of sample mixture ethanol added to the VNE Column centrifuge at 8 000 x g for 1 min Discard the flow through and the Collection Tube Combine the VNE Column with a new Collection Tube provided Add 500 ul of Wash Buffer 1 ethanol added to the VNE Column centrifuge at 8 000 x g for 1 min then discard the flow through Combine the VNE Column with the used Collection Tube Make sure that ethanol 96 100 has been added into Wash Buffer 1 when first open Add 750 ul of Wash Buffer 2 ethanol added to VNE Column centrifuge at 8 000 x g for 1 min then discard the flow through Combine the VNE Column with the used Collection Tube Make sure that ethanol 96 100 has been added into Wash Buffer 2 when first open 8 Repeat step 7 Add 750 ul of Wash Buffer 2 ethanol added to VNE Column centrifuge at 8 000 x g for 1 min then discard the flow through Combine the VNE Column with the used Collection Tube 9 Centrifuge at full speed 13 000 X g for an additional 3 min to dry the VNE column Discard the flow through and the Collection Tube Important step This step will avoid the residual liquid to inhibit the subsequent enzymatic reactions 10 Combine the VNE Column with a Elution Tube provided Add 50 ul of RNase free Water to the membrane center of the VNE Column Stand
7. umn centrifuge at 8 000 x g for 1 min then discard the flow through Combine the VNE Column with the used Collection Tube Make sure that ethanol 96 100 has been added into Wash Buffer 2 when first open 9 Repeat step 7 Add 750 ul of Wash Buffer 2 ethanol added to VNE Column centrifuge at 8 000 x g for 1 min then discard the flow through Combine the VNE Column with the used Collection Tube 10 Centrifuge at full speed 13 000 X g for an additional 3 min to dry the VNE column Discard the flow through and the Collection Tube Important step This step will avoid the residual liquid to inhibit the subsequent enzymatic reactions 11 Combine the VNE Column with a Elution Tube provided Add 50 ul of RNase free Water onto the membrane center of the VNE Column Stand VNE Column for 2 min Important step For effective elution make sure that the RNase free Water is dispensed onto the membrane center and is absorbed completely 12 Centrifuge for 2 min to elute the nucleic acid 13 Store nucleic acid at 70 C
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