Bioline ISOLATE II Plant DNA Kit Product Manual Download Page 20 | Manualshive

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PM0816V2.0

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Summary of Contents for ISOLATE II Plant DNA Kit

Page 1: ...ISOLATE II Plant DNA Kit Product Manual...

Page 2: ...Product Manual www bioline com isolate 2...

Page 3: ...pting and homogenizing starting materials 07 7 3 Lysis of plant samples 08 7 4 Buffer preparation and parameters 10 8 Protocols 11 8 1 Standard protocol for purifying plant genomic DNA 11 8 2 Support...

Page 4: ...steps with two different buffers High quality purified plant genomic DNA is then eluted in a low salt Elution Buffer or nuclease free water The DNA is ready to use for a wide variety of applications...

Page 5: ...xtremely pure genomic DNA from plant cells and tissues or filamentous fungi The DNA is suitable for a wide variety of applications e g real time PCR and DNA sequencing The preparation time is 30 min f...

Page 6: ...65o C 10 min Add 75 L Precipitation Buffer PL3 Incubate on ice 5 min Filter crude lysate 11 000 x g 2 min 1st and 2nd 11 000 x g 1 min Adjust DNA binding conditions Add 450 L Binding Buffer PB Vortex...

Page 7: ...aterial can be kept at 4 C for 24 hours but should be frozen at 20 C for longer storage 7 2 DISRUPTING AND HOMOGENIZING STARTING MATERIALS Due to the tough nature of plant tissue the lysis procedure i...

Page 8: ...mg of sample depends on both the size and genomic status DNA ploidy For example 100 mg fresh wheat containing a hexaploid genome 1 7 x 1010 bp contains 30 g DNA whereas the same amount of Arabidopsis...

Page 9: ...ant species either lysis buffer generates excellent results TABLE 1 PLANT SPECIES SUCCESSFULLY TESTED WITH ISOLATE II PLANT DNA KIT Species Organ Lysis Buffer PA1 Lysis Buffer PA2 Plant Thale cress Ar...

Page 10: ...p to 3 months For long term storage of up to 1 year divide into small aliquots and store at 20 C Elution Parameters Elute DNA using Elution Buffer PG supplied In general two consecutive elutions incre...

Page 11: ...oughly Note If sample does not resuspend easily e g due to plant powder absorbing too much buffer add more Lysis Buffer PA1 Note that the volumes of RNase A step 2 1 and Binding Buffer PB step 4 have...

Page 12: ...at the loading and centrifugation steps 6 Wash and dry silica membrane Add 400 L Wash Buffer PAW1 to the ISOLATE II Plant DNA Spin Column Centrifuge for 1 min at 11 000 x g and discard flow through No...

Page 13: ...o homogenize the sample Note If the sample cannot be easily handled because e g the sample material soaks up too much buffer additional Lysis Buffer PA1 can be added The volume of Binding Buffer PB st...

Page 14: ...ized sample into a centrifuge tube and centrifuge for 10 min at 5 000 x g Pipette 300 L of the clear supernatant into a new 1 5 mL microcentrifuge tube not supplied Proceed with step 3 filter crude ly...

Page 15: ...r Add additional lysis buffer and increase the volume of Binding Buffer PB proportionally Extraction of DNA from plant material during lysis was insufficient Increase incubation time in lysis buffer u...

Page 16: ...SSOCIATED PRODUCTS PRODUCT PACK SIZE CAT NO ISOLATE II Genomic DNA Kit 50 Preps BIO 52065 ISOLATE II RNA Plant Kit 50 Preps BIO 52077 MyTaq HS DNA Polymerase 250 Units BIO 21111 SensiFAST SYBR No ROX...

Page 17: ...Plant DNA Kit 17 NOTES...

Page 18: ...Product Manual www bioline com isolate 18...

Page 19: ...Plant DNA Kit 19...

Page 20: ...Kingdom email info uk bioline com Tel 44 0 20 8830 5300 Australia email info au bioline com Tel 61 0 2 9209 4180 Germany email info de bioline com Tel 49 0 3371 68 12 29 Singapore email info sg bioli...

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