User`s Manual
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1. bene foc Gene Foci Biotechnologies PCR Purification Kit Catalog No GF2707 User s Manual For Research Use Only In vitro Use Only PCR Purification Kit Catalog No GF2707 Catalog No Preps GF2707 50 50 GF2707 200 200 APPLICATIONS Ideal for purification of PCR product enzyme digested DNA fragment and nick translation method labeled DNA probe Also good for concentration of diluted DNA sample Kit Contents And Storage Conditions Storage PCR Purification Kit Conditions Binding Buffer BB Room Temp Wash Buffer WB Room Temp Elution Buffer EB Room Temp Binding Column EC Room Temp 2 ml Tubes Room Temp 50 preps 200 preps 50 ml 200 mi 15 ml 60 ml Add ethanol before first use 10 ml 40 ml 50 200 50 200 This kit can be stored at room temperature for up to 12 months without showing any decrease in guality and yield gt AS NOTES All buffers should be clear Lower temperature may cause precipitation If any precipitation forms warm up at 37 C water bath to dissolve before use The Gene Foci PCR purification kit should be stored at room temperature store at 4 C or 20 C may cause chemical compound precipitation in buffers Recap the bottles immediately after use to avoid unexpected oxidation evaporation and change of pH due to long term exposure to the air INTRODUCTION The Gene Foci PCR purification kit utilizes the highest qua2. temperature Not enough binding buffer BB added to PCR product Suggestion add 5x or 6x volumes of buffer BB to every 1x Low yield volume of PCR product PCR product and binding buffer BB did not mix well Suggestion gently vortex or pipette up and down several times to mix well after adding buffer BB Forget to add ethanol into buffer WB Suggestion Make sure the appropriate amount of ethanol is added into buffer WB Suboptimal elution buffer is used Suggestion Use buffer EB Poor elution to elute DNA do not use water DNA floats out of Ethanol in DNA sample Suggestion Spin dry the columns as indicated in step 8 to get rid of ethanol from the binding well while loading column agarose gel Skipped step 9 ethanol in DNA eluate Suggestion Spin dry the columns as indicated in step 8 then air dry the column for DNA is resistant to several minutes before elution enzyme digestion Silica fines in eluate Suggestion Spin the eluate at 13 000 rpm for 1 more minute use the supernatant for enzyme digestion Ordering Information To order Gene Foci products please try the following methods 1 Order online Register for an account on www Gene Foci com login and place your order using our shopping cart and secure online checking out system 2 Call our toll free number 1 888 315 9018 3 Send Email to order Gene Foci com 4 Fax your order to 1 888 959 0868 To expedite
3. the DNA ideal for PCR purification kit is between 100bp and 10 kb DNA yield drops sharply beyond this range DNA yield is correlated with the amount of DNA to begin with elution volume and the size of DNA fragment Elution buffer EB does not contain chelator EDTA thus minimizes the effect on downstream experiment such as enzyme digestion ligation Alternatively the plasmid DNA can be eluted with water However to ensure efficient elution the pH of the water must be equal or higher than 7 5 Plasmid DNA should be stored at 20 C G Spe tes oO GENE FOCI PCR PURIFICATION KIT PROTOCOL Hints gt Before start add the indicated amount of ethanol into buffer WB mix well and mark the bottle with a check For the DNA binding column EC that have been stored for more than 3 months to restore binding capacity pre treat the column with equilibrium buffer available form Gene Foci com is recommended Pre treat the binding column EC by adding 100pl eguilibrium buffer and centrifuging at 13 000rpm for 1 minute Before start it is recommended to check the PCR product with agarose gel electrophoresis to make sure the PCR reaction is successful 1 For every 100ul PCR product add 500ul binding buffer BB and mix well If the PCR product volume is smaller than 100ul adjust the volume to 100pl with ddH2O 2 Place a binding column EC into a provided 2 ml collection tube add the mixed sample from ste
4. lity silica matrix to recover DNA fragments from 100bp to 40kb free of oligonucleotides nucleotides and polymerase in yields exceeding 80 90 The binding condition of the silica matrix is adjusted by addition of a specially formulated buffer before adding the sample Following a rapid wash step DNA is eluted with EDTA free low salt and high pH elution buffer or double distilled water Purified DNA can be directly used for most downstream applications including restriction enzyme digestion ligation Sanger sequencing DNA labeling and in vitro transcription Sd HIGHLIGHTS High quality silica membranes are used to ensure the yield and consistency between different batches Unique guanidine hydrochloride and sodium iodide free binding buffer ensures high purity of DNA product gt Spe tec 2 Oo Yellow pH indicator helps monitoring the pH change in solutions and maximizing DNA binding 1to silica matrix Fast and toxic free No phenol chloroform extraction no ethanol precipitation is required ATTENTION All the steps are performed at room temperature use microcentrifuge such as Eppendorf 5415C or similar model that can handle 13 000 rpm or higher speed Binding buffer BB and equilibrium buffer contain irritating chemicals wear gloves when handling Avoid direct contact with skin eyes and clothes If contaminated rinse with large amount of water immediately The size of
5. p 1 to the column incubate at room temperature for 1 minute 3 Centrifuge at 13 000 rpm for 30 60 seconds 4 Discard the flow through place column EC back into the same tube 5 Wash column EC by adding 0 7 ml buffer WB and centrifuging at 13 000rpm for 30 seconds Make sure ethanol has been added into buffer WB 6 Discard the flow through place column EC back into the same tube 7 Repeat the wash in step 4 one more time with 0 5 ml buffer WB 8 Discard Flow through put the binding column EC back into the same 2ml centrifuge tube spin at 13 000rpm for additional 2 minutes to remove residual wash buffer Residual ethanol in buffer WB may inhibit subsequent enzymatic reactions such G se SK ec sO 4 G rec Oe 10 as restriction enzyme digestion and ligation Transfer the spin column AC into a clean 1 5 ml centrifuge tube To elute add 50 ul elution buffer buffer EB to the center of the silica membrane in the column for better yield pre warm up buffer EB to 65 70 C in a water bath incubate at room temperature for 2 minutes and centrifuge at 12 000 rpm for 1 minute For better yield re apply the eluate into the column repeat the centrifugation TROUBLE SHOOTING Problem Possible causes and suggestions Kit is stored at suboptimal temperature for example 4 C or 20 C chemicals in buffers precipitated Suggestion Always store the PCR purification kit at room
6. your order please provide the following information Customer user name Purchaser s name and detailed contact information Purchase Order Number If any Billing address Shipping address Description of the order
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