User Manual FavorPrep Gel Purification Mini Kit
Contents
1. FavorPrep Gel Purification Mini Kit User Manual Cat No FAGPK 001 50 Preps FAGPK 001 1 200 Preps For Research Use Only v 1102 Kit Contents FAGPKOO1 FAGPKOO1 1 FAGP Buffer 80 ml 160 ml X 2 Wash Buffer concentrated 15 mi 30 ml X2 Elution Buffer 5 ml 20 ml FAGP Column 50 pcs 200 pcs Collection Tube 50pcs 200pcs Add 60 ml ethanol 96 100 to Wash Buffer when first open Add 120 ml ethanol 96 100 to each Wash Buffer when first open Specification Sample size up to 400 mg agarose gel Recovery 70 85 100bp 10Kb Elution volume 40 ul Handling Time within 25 min Important Notes Interested gel slice centrifuge C centrifuge centrifuge Gel lysis FAGP Buffer Binding r Washing Wash Buffer Elution Elution Buffer Pure DNA 1 Buffers provided in this system contain irritants Wear gloves and lab coat when handling these buffer 2 Excise the extra agarose gel to minimize the size of the gel The maximum size of the gel slice is 400 mg 3 Add the indicated volume of ethanol 96 100 to Wash Buffer when first open 4 All centrifuge steps are done at full speed 13 000 rpm or 17 900 x g in a microc entrifuge Genernal Protocol Please Read Important Notes Before Starting The Following steps HINT Prepare a 55 C dry bath or water bath for step 4 1 Excise the agarose gel containing relevant DNA fragments with a clean scalpel2. e Remove the extra agarose gel to minimize the size of the gel slice The optimal size of the gel slice is around 200 mg 2 Determine the weight of the gel slice and transfer it to a clean tube not provided eThe maximum size of the gel slice is 400 mg 3 Add 3 volumes of FAGP Buffer For example Add 600 ul of FAGP Buffer to 200 mg of gel slice 4 Incubate at 55 C for 10 15 min and vortex the tube every 2 3 min until the gel slice dissolved completely e During incubation interval vortex can accelerate the gel dissolved Important Make sure that the gel slice has been dissolved completely before proceed the next step 5 Vortex the sample mixture for 5 seconds Cool down the sample mixture and place a FAGP column into a Collection Tube 6 Transfer 800 pl of the sample mixture to the FAGP Column Centrifuge for 30 seconds discard the flow through and place the FAGP column back to the Collection Tube elf the sample mixture is more than 800 ul repeat the step 6 for the rest sample mixture 7 Add 750 ul of Wash Buffer ethanol added to the FAGP Column Centrifuge for 30 seconds discard the flow through and place the FAGP column back to the Collection Tube e Make sure that ethanol 96 100 has been added to Wash Buffer when first open elf the downstream application is very salt sensitive stand the FAPG Column for 3 min after adding Wash Buffer before centrifuging 8 Centrifuge for an additional 2 min
3. non specific DNA fragment e Contaminated scalpel Using a new or clean scalpel e DNA fragment is denatured Incubate eluted DNA at 95 C for 2 min then cool down slowly to reanneal denatured DNA Poor performance in the downstream applications e Salt residue remains in eluted DNA fragment Wash the column twice with Wash Buffer e Ethanol residue remains in eluted DNA fragment Do discard the flow through after washing with Wash Buffer and centrifuge for an additional 3 min
4. to dry the FAGP column eImportant Step This step will avoid the residual liquid to inhibit subsequent enzymatic reactions 9 Place the FAGP Column into a new 1 5 ml microcentrifuge tube not provided 10 Add 40 ul of Elution Buffer or ddH20 pH7 0 8 5 to the membrane center of the FAGP Column Stand the FAGP Column for 1 min e Important step For effective elution make sure that the elution solution is dispensed onto the membrane center and is absorbed completely Important Do not elute the DNA using less than suggested volume 40uyl It will lower the final yield 11 Centrifuge for 1 min to elute the DNA Troubleshooting e Agarose gel of high percentage gt 2 is used Add 6 volumes of FAGP Buffer to 1 volume of the gel slice e The size of the gel slice is too large If the gel slice is more than 400 mg separate it into multiple tubes Low or none recovery of DNA fragment e The column is loaded with too much agarose gel The maximum volume of the gel slice is 400 mg per column Make sure that the gel slice has been dissolved completely before proceed the next step e Elution of DNA fragment is not efficient Make sure that the volume of elution solution is 2 40 ul Make sure that the elution solution has been completely absorbed by the membrane before centrifugation e The size of DNA fragment is larger than 5 Kb Preheat the elution solution to 60 C before use Eluted DNA contains
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