柱式质粒中量抽提试剂盒(50～100mL)

产品货号：

GS2251

中文名称：

柱式质粒中量抽提试剂盒(50～100mL)

英文名称：

SD-200 Spin Column Plasmid DNA Mid-Preps Kit(50-100mL)

产品规格：

10T|20T

发货周期：

1～3天

产品价格：

询价

This kit provides a simple and efficient method for large quantity of plasmid DNA purification.DNA fragments are selectively adsorbed in silica gel-based column and other impurities such as proteins,salts,oligos (＜40-mer) and nucleotides are washed away.Kit contains a unique embedded in a 25mL spin column for binding up to 200mg of plasmid DNA.Purified Plasmid DNA can be used for any downstream applications such as sequencing,restriction reactions,labeling,transformation,PCR and Southern-blotting.

Fast.Entire procedure takes 30～40minutes.
Preparation of high quality plasmid DNA from culture.Purified DNA can be used in any downstream applications such as sequencing,transformation,restriction enzymatic digestion,and transfections.
High yields (80～90%) and Reproducible.
No phenol/chloroform extraction;No CsCl centrifugation;No ethanol precipitation

components	10T	20T
Solution I	55mL	110mL
Solution II^a	55mL	110mL
Solution III	75mL	2′75mL
Wash Solution^b	24mL	48mL
Elution Buffer^c	15mL	30mL
RNase A (10mg/mL)^d	0.8mL	1.6mL
SPIN200 Column	10	20
50mL Collection Tube	10	20

注：

Solution II may form a precipitate upon storage.If necessary,dissolve the precipitate by warming at 370C.
Before use,add 96mL of 100% of ethanol to 24mL Wash Solution for 10T,add 192mL of 100% ethanol to 48mL Wash
Solution for 20T.If volume of Wash Solution is not sufficient as 24mL or 48mL due to leaking during transportation,it is
necessary to re-measure its volume,and determine how many ml of ethanol should be added (volume of added ethanol:volume of Wash
Solution=4:1).
Elution Buffer is 2.0mM Tris-HCl pH8.5.the resulting yield is add up to 120% than TE buffer pH8.0 or water can be used,
Before use add RNase A to Solution I.Then Solution I Should be stored at 4℃ for frequent use,or stored at -20℃ if not use for a long period.

Bacterial cultures are lysed and the lysates are cleared by routine methods (Solution I,II,III).Plasmid DNA from the cleared lysates is selectively bound to the silica-based membrane in the column and impurities are washed away.Pure DNA is eluted in low-salt buffer or water.

Add 50～100mL overnight culture (OD₆₀₀=2.5～5) to a approprite centrifuge tube and centrifuge at 5000rpm for 10minutes.Drain the liquid completely.
Note:can not be loaded excess bacterials,or else can not be lysed fully
Add 5mL of Solution I to the pellet,mix gently and keep on ice for 2minute.
Note:the bacterials must be suspended out and out,or else can not be lysed fully
Add 5mL of Solution II to the mixture,mix gently by inverting the tube 4-6 times and then keep at RT for 2minutes.
Note:To prevent contamination from genomic DNA,do not vortex drastically.and the whole lysis time can not exceed 5minutes
Add 7mL of Solution III,and mix gently.Incubate at RT for 2minutes.
Spin at 10000rpm for 10minutes.
Place column into a 50mL collection tube.Transfer the above supernatant (step 5) into the column,Stand at RT for 5minutes.Spin at 6000rpm for 3-5minutes.
Discard the flow-through in the tube.Add 5mL of Wash Solution to the column,and spin at 6000rpm for 3-5minutes.
Repeat wash procedure in step 7.
Discard the flow-through in the collection tube.Spin at 6000rpm for additional 5minutes to remove residual Wash Solution.
Note:In order to gain high plasmid DNA elution efficiency,keep the SPIN200 column at room temperature for 10minutes or in dryer at 50℃ for 5minutes,thus the residual ethanol is volatile thoroughly
Transfer the column to a clean 50mL microfuge tube.Add 500μL of Elution Buffer into the center part of the membrane of the column and incubate at 37～50℃ for 2minutes.Spin at 6000rpm for 2minutes.Store DNA in freezer at–20℃.
Note:In order to gain high plasmid DNA elution efficiency,the elution buffer or ddH2O can be heated to 60℃ and then apply them to the SPIN200 column.
Note:It is important to add the Elution Buffer into the center part.Pre-warm Elution Buffer at 55～80℃ could increase elution efficiency.Two times elution is recommended.

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