A protocol for DNA extraction in barley (Brandes, in prep.) was slightly modified for DNA extraction in caraway. 192 plants per each F 1 population (n = 7) were grown in greenhouse in 96 pot plates. The youngest developing leaf per plant (BBCH = 15-18 (Hack et al. 1992 )) was sampled. Leaves were collected in racked collection microtubes (Cat. no. 19560, Qiagen) . Two 4 mm steel beads per sample were added. Samples were frozen with liquid nitrogen. Samples were ground using mixer mill MM 300 (Retsch). Samples were incubated at 65 °C for 45 min after adding 600 ll extraction buffer (1 M guanidine thiocyanate, 2 M NaCl, 30 mM NaAc) per sample. Samples were spun using Rotina 420 centrifuge (Hettich) for 30 min at 4000 rpm. 400 ll of liquid per sample were transferred to new collection microtubes and 2.5 ll RNase A (5 mg/ml) was added. Samples were incubated for 30 min at 35 °C. DNA was precipitated, adding 300 ll propan-2-ol per sample. Samples were spun using Rotina 420 centrifuge (Hettich) for 30 min at 4000 rpm. DNA pellets were washed in two steps, adding 600 ll per solution and sample (wash solution 1: 76% ethanol, 200 mM NaAc; wash solution 2: 76% ethanol, 10 mM NaAc). Dried DNA was dissolved in 600 ll TE-buffer (10 mM Tris/HCl, 1 mM EDTA) per sample. DNA was quantified using nanodrop 8000 spectrophometer (Thermo Fisher Scientific).
Rotina 420 centrifuge
Manufactured by Hettich
Sourced in Germany
The Rotina 420 is a general-purpose laboratory centrifuge designed for routine applications. It features a maximum speed of 4,500 rpm and a maximum RCF of 3,020 x g. The centrifuge can accommodate various rotor types and sample volumes.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using rotina 420 centrifuge
1
DNA Extraction Protocol for Caraway
2
DNA Extraction Protocol for Caraway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A protocol for DNA extraction in barley (Brandes, in prep.) was slightly modified for DNA extraction in caraway. 192 plants per each F 1 population (n = 7) were grown in greenhouse in 96 pot plates. The youngest developing leaf per plant (BBCH = 15-18 (Hack et al. 1992 )) was sampled. Leaves were collected in racked collection microtubes (Cat. no. 19560, Qiagen) . Two 4 mm steel beads per sample were added. Samples were frozen with liquid nitrogen. Samples were ground using mixer mill MM 300 (Retsch). Samples were incubated at 65 °C for 45 min after adding 600 ll extraction buffer (1 M guanidine thiocyanate, 2 M NaCl, 30 mM NaAc) per sample. Samples were spun using Rotina 420 centrifuge (Hettich) for 30 min at 4000 rpm. 400 ll of liquid per sample were transferred to new collection microtubes and 2.5 ll RNase A (5 mg/ml) was added. Samples were incubated for 30 min at 35 °C. DNA was precipitated, adding 300 ll propan-2-ol per sample. Samples were spun using Rotina 420 centrifuge (Hettich) for 30 min at 4000 rpm. DNA pellets were washed in two steps, adding 600 ll per solution and sample (wash solution 1: 76% ethanol, 200 mM NaAc; wash solution 2: 76% ethanol, 10 mM NaAc). Dried DNA was dissolved in 600 ll TE-buffer (10 mM Tris/HCl, 1 mM EDTA) per sample. DNA was quantified using nanodrop 8000 spectrophometer (Thermo Fisher Scientific).
3
Honey Fingerprinting with HPLC-UV
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Honey samples analyzed with the universal non-targeted HPLC-UV fingerprinting method were prepared as follows: approximately 1 g of honey was weighed in a 15 mL PTFE centrifuge tube (Serviquimia, Barcelona, Spain) and dissolved in 10 mL of Milli-Q water by mixing the contents using a VibraMix Vortex from OVAN (Barcelona, Spain). Then the samples were centrifuged for 5 min at 3500× g rpm (Rotina 420 Centrifuge from Hettich, Tuttlingen, Germany) to separate any non-soluble particles (that may include bee bread, pollen, proteins, etc.). The obtained extracts were then diluted with methanol in a 1:1 ratio, filtered through 0.45 µm syringe membrane filters (FILTER-LAB, Barcelona, Spain), and kept at 4 °C until analysis. Honey samples were randomly analyzed. In the case of crystallized honeys (which is a normal state of natural raw honeys), they were first introduced in a water bath at 45 °C until they melted. After their homogenization and cooling, they were treated following the same procedure.
50 µL of each diluted extract were mixed to prepare a quality control (QC) sample that was used to evaluate the repeatability and the robustness of the proposed non-targeted HPLC-UV fingerprinting methodology and that the chemometric results were not affected by instrumental drifts.
50 µL of each diluted extract were mixed to prepare a quality control (QC) sample that was used to evaluate the repeatability and the robustness of the proposed non-targeted HPLC-UV fingerprinting methodology and that the chemometric results were not affected by instrumental drifts.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!