Manual
Contents
1. 50 Buffer BL Buffer GC DNA Wash Buffer Elution Buffer Mini Columns User Manual Before Starting Prepare all components and get all necessary materials ready by examining this instruction booklet and become familiar with each steps Important Add 8ml XG3511 XG3514 00 or 60ml XG3511 XG3514 01 100 ethanol to DNA Wash Buffer before use A gelslice of 100 mg equals to a volume of 100 ul Buffer GC may form precipitates under cool ambient condition Warm up the buffer at 37 C to dissolve before use Buffer BL precipitates below room temperature It is critical to warm up the buffer at 50 Cto dissolve the precipitates before use Keep the cap tightly closed for Buffer BL after use Pre warm aliquots of Elution Buffer or ddH O at 55 60 C waterbath Safety Information Buffer GC contains acidic acid and chaotropic salts which may form reactive compounds when combines with bleach Do not add bleach or acidic solutions directly tothe preparation waste Performall stepsincluding centrifugation at room temperature o 3 DNA Gel PCR Purification Spin Protocol Note Fresh TAE buffer as running buffer is recommended Reusing running buffer will result the increase ofthe pHand then reduce yield 1 Add 400ul of Buffer BL into the spin column incubate at room temperature for 2 minute centrifuge for 2 minute at 12 000 rpm and discard the flow through The column is ready and will work well for binding DNA 2 Forcycl2. L z enomics Table of Contents Odi 0 ne O 02 OV A AE E 02 Sera ae BE A AR L 02 KiE COMINS non TA AE A AAA oc cco encore eee 03 BEOS S _AaE W MR 03 SEE LUO NEC hi A A M A 03 DNA el PGPEUICaton Spin lol NU 04 DNA Gel PCR Purification Vaccum Spin Protocol iiiisesnrennronnnnnnoonnmu 05 MoubleshooingiGuide S fe Wi OU 07 BEC SOC WalentyP gt SA u 08 01 Introduction This fast and reliable kit is designed to recover DNA from agarose gels and purify DNA fragments from PCR RFLP phosphorylation labeling ligation hybridization and other enzymatic reactions DNA fragments from 100 bp to 20 kb can be purified using the mini column with over 80 90 recovery Overview If using the DNA Gel PCR Purification Mini Kit for the first time please read this booklet to become familiar with the procedures Samples are homogenized and lysed in a high salt buffer The DNA is bound to the column while proteins and other impurities are removed by wash buffer The purified DNA or PCR products is suitable for downstream applications such as endonuclease digestion thermal cycle amplification and hybridization technigues Storage and Stability All components can be stored at room temperature All kit components are stable up to 12months 02 DNA Gel PCR Purification xcelris genomics M i n i Kit An Abellon company XcelGen Kit Contents Product XG3511 00 XG3511 01 XG3514 00 XG3514 01 Preps
3. dd ethanol to DNA Wash Buffer Ethanol was not completely removed from the column following wash step Agarose gel doesn t melt completely xcelris Abellon Suggestion 1 Determinethe volume of Buffer GC to be used correctly as instructed 2 Make sure to set the water bath to 55 60 C to allow gel to melt completely Add more Buffer GC if necessary 3 Usefresh electrophoresis buffer 4 Incubate the column after adding ddH 0 or Elution Buffer at 60 C for 15 min before elution Add absolute ethanol to DNA Wash Buffer as instructed before use After the wash step centrifuge the empty column with the lid open at top speed for 1 3 min Repeat once Make sure to melt the gel at 55 60 C before loading the sample to DNA column Limited Use and Warranty This productis intended for in vitro research use only Not for use in human This product is warranted to perform as described in its labeling and in XcelGen s literature when used in accordance with instructions No other warranties of any kind express or implied including without limitation implied warranties of merchantability or fitness for a particular purpose are provided by XcelGen XcelGen s sole obligation and purchaser s exclusive remedy for breach of this warranty shall be at the option of XcelGen to replace the products XcelGen shall have no liability for any direct indirect conseguential or incidental damage arising out of the use the r
4. e pure PCR reaction Add 3 volumes of Buffer GC to 1 volume of the PCR reaction and mix completely by vortexing Briefly spin the tube to collect any drops from the inside wall and tube lid For agarose gel Excise the DNA fragment from the agarose gel and weigh it in a 1 5 ml microfuge tube Add 3 volume of Buffer GC to 1 volume of gel to the 1 5 ml microfuge tube and incubate the mixture at 55 60 C for 8 minute Mix the tube by tapping the bottom every 2 minute till the gel has melted completely Cool the tube to room temperature Add 1 volume of isopropanol 3 Transfer up to 700pl DNA Buffer GC mixture to a spin column with a collection tube Centrifuge at 13 000g for 1 minute at room temperature Discard the flow through and put the column back to the collection tube Repeat this step to process the remaining solution 4 Add 500pl of Buffer GC into the DNA Mini Column Centrifuge at 13 000g for 1 minute at room temperature to wash the DNA Mini Column 5 Add 650ul DNA Wash Buffer to the column and centrifuge at 13 000g for 1 minute at room temperature Discard the flow through and insert the column with the lid open back to the collection tube Repeat step 5 Note Ensure that ethanol has been added to DNA Wash Buffer as instructed 6 Centrifugethe empty DNA column with the lid open at 13 000g for 2 minute to dry the ethanol residue in the matrix Note The residual ethanol will be removed more efficiently with the column l
5. esults of use or the inability to use it product For technology support or learn more product information please visit our website at www xcelrisgenomics com 08 XcelGen Quality Kits made by Xperts Plasmid DNA Isolation Kits Genomic DNA Extraction Kits RNA Extraction Kits Polymerase DNA Ladders DNA Markers Premix Tag dNTP s RAPD kits Agarose Glycerol Tms NA Stabilizers 8 RNA Protectant solutions PrimeX Oligo Synthesis amp Purification Services 10 nmole 25 nmole 50 nmole 100 nmole 200 nmole 1000 nmole NGS Services Denovo Genome Sequencing Whole Genome Resequencing GBS RAD Sequencing Exome Sequencing Amplicon Sequencing Whole Transcriptome Analysis RNA Sequencing Small RNA Sequencing Metagenomics Metatranscriptomics ChIP Sequencing Mitochondrial Sequencing Next Generation Genomic Services on Illumina MiSeq Genotyping by Sequencing Tilling Ecotilling using NGS Genome Database development Services NGS Bioinformatics e In silico Primer Design Microarray Analysis Metagenomics Physical Genetic and QTL mapping Assembly and annotation of prokaryotic and eukaryotic genome Genome Mapping and SNP discovery Transcriptome discovery and analysis sRNA analysis and discovery XcelSeq Sanger Sequencing Servi Plasmid PCR Sequencing Services r E coli Culture Sequencing Services Primer Walk Sequencing Services Microbial Identification Ser
6. id open during centrifugation 7 Place the column into a clean 1 5 ml microfuge tube and add 30 50 ul pre warmed 60 C Elution Buffer or ddH O to the center of the column Incubate at room temperature for 1 minute Centrifuge at 13 000g for 1 minute to elute the DNA Reload the eluted DNA solution to the column fora second elution 04 Note Pre warm elution buffer or ddH 0 at 60 C and incubate the column at 60 C for 5 minute after adding Elution Buffer or ddH 0 willincrease the DNA yield Note For fragment larger than 8 kb incubate the column at 60 C for 15 minute after adding Elution Buffer or ddH 0willincrease the DNA yield Note The first elution normally yields 60 70 of the DNA bound Reload the eluted DNA solution to the column for a second elution will yield another 20 ofthe DNA that makes the total yield up to 90 DNA Gel PCR Purification Vaccum Spin Protocol tle 05 Follow the instruction described on step 1 amp 2 on page 4 Briefly spin the tube to collect any drops from the inside wall and tube lid Prepare the vacuum manifold according to manufacturer s instructions Attach the spin column to the manifold Load the Gel or PCR reaction Buffer GC solution to a spin column that is attached to the manifold Turn on the vacuum to let the solution pass through the column Wash the column by adding 650ul DNA Wash Buffer Repeat step 4 Vacuum the column for 1 minute Put the column with the l
7. id open in a collection tube and spin at top speed for 3 minute Note The residual ethanol will be removed more efficiently with the column lid open during centrifugation Put the column to a clean 1 5 ml microfuge tube and add 30 50pl Elution Buffer or ddH O to the column Incubate at room temperature for 2 minute Centrifuge the tube at 13 000g for 1 minute to elute DNA Note Pre warm Elution Buffer or ddH 0 at 60 C and incubate the column at 60 C for 5 minute after adding elution buffer or ddH 0 willincrease the DNA yield Note The first elution normally yields 60 70 of the DNA bound Reload the eluted DNA solution to the column for a second elution will yield another 20 of the DNA that makes the total yield up to 90 on DNA Gel PCR Purification xcelris celGen Mini kit ee Abellon Before Gel purification After Gel purification 100 bp 1kb 100bp 500bp 3kb 1kb ladder ladder ladder Product Product ladder lt 3kb lt 3kb E 500 bp gt Before Gel purification After Gel purification 20 kb gt gt Genomic DNA Gel Purified genomic DNA 06 Gen DNA Gel PCR Purification Mini Kit Troubleshooting Guide Problems Low Yield No DNA yield DNA sample floats out of well while loading agarose gel 07 Possible Reasons 1 Not enough Buffer GC 2 Agarose gel doesn t melt completely 3 Reused electrophoresis Buffer with increased pH 4 Fragment gt 10 kb Forgot to a
8. vice Multilocus Sequence Typing Customised Services SNP Genotyping by SNaPshot Assay Microsatellite Genotyping Golden Gate Assays and Arrays Gene Expression on Real Time PCR Gene expression on Agilent Microarray Affymetix Library construction Xcelris Labs Limited e xce rl S Old Premchand Nagar Road Opp Satyagrah Chhavani Bodakdev Ahmedabad 380015 India Tel 91 79 66197777 Fax 91 79 66309341 genomics Website www xcelrisgenomics com An Abellon Company E mail bdgenomics xcelrislabs com
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