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MagListo 5M Скачать руководство пользователя страница 21

 

 

18 

www.bioneer.com

 

BQ-042-101-05 

Revision : 2 (2016-07-04)

 

 

d. DNA Extraction from Gram-Negative Bacteria for Mini/Midi/Maxi 

 

1.

 

(Cell  collection)

  Collect  the  bacteria

  cells  (~1x10

9

  (mini)/  ~5x10

9

  (midi)/  ~1x10

10

  (maxi))  by 

centrifugation at 6000 x g (> 8,000 rpm in a microcentrifuge) for 10 min (or >3,500 rpm for 20 min). 

And completely remove the media by pipetting. 

 

2.

 

(Resuspension of cell pellet: 2-3)

 Add 180 μl (mini)/ 1.8 ml (midi)/ 3.6 ml (maxi) of Buffer  ⓛ  (Lysis) 

to the collected cell pellet and completely resuspend by vortex mixer or pipetting. 

 

3.

 

Transfer the resuspended cell pellet to each specific tube format. 

a. (Mini) Transfer the resuspended pellet to a 1.5 ml or 2 ml tube. 

b. (Midi) Transfer the resuspended pellet to a 15 ml tube. 

c. (Maxi) Transfer the resuspended pellet to a 50 ml tube. 

 

4.

 

Go to step 3 of “DNA Extraction from Animal Tissue” in page 14 and continue the extraction process.

 

Содержание 5M

Страница 1: ......

Страница 2: ...Bioneer Corporation 8 11 Munpyeongseo ro Daedeok gu Daejeon 34302 Republic of Korea Tel 82 42 930 8777 Fax 82 42 930 8688 Email sales bioneer co kr www bioneer com...

Страница 3: ...l Amount of Sample 4 X Principle 4 XI Materials and Equipment Needed But Not Provided 5 XII Protocols 6 Before You Begin 6 DNA Extraction from Whole Blood 6 DNA Extraction from Cultured Cell 10 DNA Ex...

Страница 4: ...e rapid extraction No costly capital instrument to instruments except MagListoTM Magnetic Separation Rack Applications Gene Cloning PCR Real Time PCR Southern Blotting SNP genotyping II Kit Components...

Страница 5: ...ly recombinant samples including tubes tips and materials should be processed and disposed of according to applicable and appropriate regulations of the municipality government in which this product i...

Страница 6: ...o your needs If you have any questions or would like to find out more information about MagListo products please contact us We look forward to hearing from you Technical Support For all technical ques...

Страница 7: ...mer such as poly dA poly dT or gDNA or RNA should be added to the starting material Ensure that the carrier DNA does not interfere with your downstream application Carrier RNA can be removed later by...

Страница 8: ...aline PBS 6 Blow dryer or heat gun 7 MagListoTM Magnetic Separation Rack Magnetic Separation Rack Choice Tube MagListoTM Magnetic Separation Rack 1 ml tube with 8 cap strip MagListoTM 8Ch Magnetic Sep...

Страница 9: ...di 200 l maxi of proteinase K see Before you begin to each specific tube format a Mini Add proteinase K to a 1 5 ml or 2 ml tube b Midi Add proteinase K to a 15 ml tube c Maxi Add proteinase K to a 50...

Страница 10: ...htly bind to the magnet Attachment Combine the magnet plate to the stand 8 Without removing the tube from MagListoTM carefully pour the supernatant out and completely remove the remaining supernatant...

Страница 11: ...tep 9 11 by adding 700 l mini 5 ml midi 10 ml maxi of Buffer 2 nd Washing for additional washing 14 3 rd washing Repeat the above step 9 11 by adding 700 l mini 5 ml midi 10 ml maxi of absolute ethano...

Страница 12: ...etic nano beads Do not reuse the beads Summary of Reagents Volume in Each Step of DNA Extraction from Whole Blood Step Buffer Mini Midi Maxi Page Blood PBS 200 l 2 ml 4 ml P 6 Lysis Buffer Binding 200...

Страница 13: ...ro 20 l mini 100 l midi 200 l maxi of proteinase K see Before you begin to the tube 4 If RNA free genomic DNA is required add up to 2 l micro 10 l mini 75 l midi 150 l maxi of RNase A see Before you b...

Страница 14: ...g a paper towel by blotting Discard solution Discard solution by inverting the MagListo rack The silicone immobilizer inside the stand holds the tubes from falling in an upside down position When disc...

Страница 15: ...for 1 min micro mini 3 min midi 5 min maxi 3 cm away from the top of the tube Note Without using a heat gun or a blow dryer place the rack lying down in a dry oven at 60 for 10 min micro mini 20 min...

Страница 16: ...10 7 P 10 Lysis Buffer Binding 100 l 200 l 1 ml 1 ml P 10 DNA Precipitation Absolute Ethanol 200 l 400 l 2 ml 2 ml P 10 Bead Binding Magnetic Nano Bead DNA 100 l 100 l 500 l 1 ml P 10 1 st Washing Bu...

Страница 17: ...100 l midi 200 l maxi of proteinase K see Before you begin to the tube and mix thoroughly using a vortex mixer 4 If RNA free genomic DNA is required add up to 10 l mini 75 l midi 150 l maxi of RNase A...

Страница 18: ...out and completely remove the remaining supernatant using a paper towel by blotting Discard solution Discard solution by inverting the MagListo rack The silicone immobilizer inside the stand holds th...

Страница 19: ...e beads with the tube open and use a heat gun or a blow dryer for 1 min mini 3 min midi 5 min maxi 3 cm away from the top of the tube Note Without using a heat gun or a blow dryer place the rack lying...

Страница 20: ...Lysis Buffer Lysis 180 l 1 8 ml 3 6 ml P 14 Buffer Binding 200 l 2 ml 4 ml P 14 DNA Precipitation Absolute Ethanol 400 l 4 ml 8 ml P 14 Bead Binding Magnetic Nano Bead DNA 100 l 500 l 1 ml P 14 1 st...

Страница 21: ...a by pipetting 2 Resuspension of cell pellet 2 3 Add 180 l mini 1 8 ml midi 3 6 ml maxi of Buffer Lysis to the collected cell pellet and completely resuspend by vortex mixer or pipetting 3 Transfer th...

Страница 22: ...3 Transfer the resuspended cell pellet to each specific tube format a Mini Transfer the resuspended pellet to a 1 5 ml or 2 ml tube b Midi Transfer the resuspended pellet to a 15 ml tube c Maxi Transf...

Страница 23: ...thanol to the eluate and mix well by vortex mixer 5 DNA binding with Magnetic Nano Bead 5 7 Add 100 l mini 500 l midi 1 ml maxi of Magnetic Nano Bead solution to the tube and mix thoroughly using a vo...

Страница 24: ...MagListo rack Add 700 l mini 5 ml midi 10 ml maxi of Buffer 2 nd Washing to the tube Close the cap and mix by vortex mixer until beads are fully resuspended Detachment Push up the magnet plate gently...

Страница 25: ...rity from turbid to clear indicating that protein digestion has occurred Extend the incubation time if unlysed tissue is still present The amount of time for complete lysis varies depending on the typ...

Страница 26: ...e eluate refer to DNA Clean Up protocol in page 20 for details Aggregation of Magnetic Nano Bead There is an excess amount of sample to extract DNA Appropriate amount of starting material see product...

Страница 27: ...R Purification Kit 8 reactions mini 1 kit K 3609 MagListoTM 5M PCR Purification Kit 100 reactions mini 1 kit K 3610 MagListoTM 5M Cell Total RNA Extraction Kit 8 reactions mini 1 kit K 3611 MagListoTM...

Страница 28: ...umber Contains sufficient for n tests USE BY Consult Instruction For Use Batch code Caution consult accompanying documents Temperature Limitation In Vitro Diagnostics Medical Device Manufacturer Cauti...

Страница 29: ......

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