Spin column based genomic DNA purification from yeast
For general laboratory use.
Shipping: shipped at ambient temperature
Storage Conditions: store at ambient temperature
Shelf Life: 12 months
The obtained DNA is suitable for a variety of applications, including real-time PCR, southern blot analysis, genotyping and discovery or validation of SNP/SSR markers.
Before start, add the following components as indicated on the respective bottle/tube:
The spin column based Yeast DNA Preparation Kit is designed for rapid and high purity isolation of genomic DNA from yeast cells. The spin column based method completely removes PCR inhibitors such as divalent cations and proteins resulting in a high purity preparation of genomic DNA. There is no use of phenol or chloroform, handling is safe and does not produce any harmful waste.
Column based genomic DNA purification kits yield up to 30 μg DNA sized from 200 bp to 50 kb per preparation.
To be provided by you:
96-99 % Ethanol
Double distilled water
Microtubes 1.5 or 2.0 ml
For S pack (50 preps): Before start, add 500 μl dd-water to the Proteinase K tube, 70 μl Lyticase Buffer to the Lyticase tube, 150 μl dd-water to the RNase A tube and 48 ml 96-99 % Ethanol (not included in the kit) to the Washing Buffer bottle.
For L pack (5 x 50 preps): Before start, add 500 μl dd-water to each Proteinase K tube, 70 μl Lyticase Buffer to each Lyticase tube, 150 μl dd-water to each RNase A tube and 120 ml 96-99 % Ethanol (not included in the kit) to each Washing Buffer bottle.
|Resuspension Buffer||16 ml||5 x 16 ml|
|175 units||5 x 175 units|
|Lyticase Buffer||70 μl||5 x 70 μl|
|Lysis Buffer||16 ml||5 x 16 ml|
|Binding Buffer||16 ml||5 x 16 ml|
|7.5 mg||5 x 7.5 mg|
|5 mg||5 x 5 mg|
|Activation Buffer||6 ml||5 x 6 ml|
|Washing Buffer||add 120 ml Ethanol to each bottle (final volume 150 ml each)||add 120 ml Ethanol to each bottle (final volume 150 ml each)|
|Elution Buffer||5 ml||5 x 5 ml|
It is essential to use the correct amount of starting material in order to obtain optimal DNA yield and purify. A maximum amount of
1 - 2 x 107 yeast cells can generally be processed. Overnight cultured yeast cells can be processed. Cell pellets can be stored at -70 °C for several months.
1 Cell Lysis:
B Column Activation [optional]:
C Column Loading:
D Primary Washing:
E Secondary Washing:
F Elution of DNA:
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