[EasyTools] PCR Mixture Calculator

PCR Mixture Calculator

Sample Count:

Repeat Count:

For 10 pmol stock primer For 100 pmol stock primer

Calculation Result:

Typically, the cDNA concentration used in qPCR falls within the range of 10 ng/μL to 100 ng/μL. This range is commonly used in various experiments, but it may be adjusted based on the experiment's objectives and sample types.

This calculator is based on a cDNA volume of 1ul. PCR mixture 19 ul + cDNA 1 ul

Mouse Genotyping

Mouse Genotyping protocol

Step 1: Gather Materials and Reagents

• Make sure you have all the necessary materials and reagents ready for the genotyping process. This includes the tissue samples (tail snips, ear punches, or other tissues), Extraction Reagent, Stabilization Buffer, PCR reagents, and agarose gel.

• AccuStart™ II Mouse Genotyping Kit (Cat. No. 95135-100; Size: 100 reactions; Quantabio)

Step 2: Prepare Tissue Samples

• Depending on the type of tissue samples you have (tail snips, ear punches, or other tissues), determine the appropriate volume of Extraction Reagent needed. Refer to the table provided earlier for guidance on sample sizes and Extraction Reagent volumes.

• Ensure that tissue samples are small and completely submerged in Extraction Reagent.

Sample Type	Sample Size	Extraction Reagent Volume	Comments
Tail Snips	2 mm	100 μL	Fresh or frozen tail snips can be used. Use proportionally more Extraction Reagent with tail snips larger than 2 mm. Samples will appear to remain intact and will not dissolve in Extraction Reagent after heating.
Ear Punches	2 mm	50 μL	Ensure that the ear punches are completely submerged in Extraction Reagent.
Other Tissues	5 mg	100 μL	Tissue samples should be small and completely submerged in Extraction Reagent.

Step 3: DNA Extraction

• Add the tissue samples to the appropriate volume of Extraction Reagent as calculated in Step 2.

• Heat the samples to 95°C for 30 minutes. Note that the tissue samples will not dissolve and will appear intact after incubation.

• Cool the samples to room temperature.

• Add an equal volume of Stabilization Buffer to the cooled samples. This serves as a safe stopping point.

• Extracts can be stored at 4°C for several weeks or at -20°C for several months.

Step 4: PCR Reaction Setup

• Use up to 2.5 μL of the DNA extract in a 25 μL PCR reaction. Depending on the sample size and extraction conditions, you may need to dilute the extract in water or TE buffer for optimal PCR results.

• Follow your standard PCR reaction setup protocols for amplifying the DNA regions of interest.

Step 5: Gel Electrophoresis

• Load 5 or 10 μL (or as appropriate) of the PCR products on an agarose gel.

• Perform gel electrophoresis to visualize and confirm the presence of the desired DNA fragments.

Additional Notes:

The protocol can be optimized for specific applications by adjusting incubation times or omitting the addition of Stabilization Buffer if needed.

It's recommended to use PCR tubes or multi-well plates and incubate in a thermal cycler with a heated lid to prevent sample condensation.

[EasyTools] Protein Concentration Calculation

Protein Concentration Calculation from BCA assay

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How to Use

1. Enter the concentration of each sample in the "Sample Concentrations (ug/mL)" section. You can leave some fields empty.

2. Input the common dilution factor and final sample volume.

3. Click the "Calculate Dilution" button to perform the calculation.

4. View the calculation results in the table. The sample with the lowest corrected concentration will be highlighted.

5. The required loading volumes for 50ug, 30ug, and 10ug of protein will be displayed.

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Sample 1 Concentration (ug/mL):

Sample 2 Concentration (ug/mL):

Sample 3 Concentration (ug/mL):

Sample 4 Concentration (ug/mL):

Sample 5 Concentration (ug/mL):

Sample 6 Concentration (ug/mL):

Sample 7 Concentration (ug/mL):

Sample 8 Concentration (ug/mL):

Sample 9 Concentration (ug/mL):

Sample 10 Concentration (ug/mL):

Common Dilution Factor:

Final Sample Volume (uL):

4x Loading Buffer: 0 uL

Dilution Calculation Result:

Sample	Original Concentration (ug/mL)	Protein (ug/uL)	Lysate (uL)	Lysis Buffer (uL)

Loading Volume (uL):

BCA Protein Assay Protocol

Pierce™ BCA Protein Assay Protocol

Materials and Equipment:

• Pierce™ BCA Protein Assay Kit (A55864)
• Sample (protein sample)
• BSA (Bovine Serum Albumin for creating a standard curve)
• Microplate (96 wells)
• Multi-pipette (for dispensing samples and standards)
• Spectrophotometer or plate reader

Steps:

1. Add your protein samples to each well of the microplate. Add 25-200 μg of protein to each sample well.

2. Dilute the standard solution to various concentrations, typically 0, 7.8, 15.6, 31.25, 62.5, 125, 250, 500 μg/mL, etc.

3. Prepare the Pierce™ BCA Reagent mixture: Mix 50 parts of Pierce™ BCA Reagent Solution A with 1 part of Pierce™ BCA Reagent Solution B. For example, mix 50 μL of Solution A with 1 μL of Solution B to create the working Pierce™ BCA Reagent solution. (A:B=50:1)

4. For each standard concentration and sample, add the Pierce™ BCA Reagent and mix well. Incubate the plate at 37°C for 30 minutes.

5. After the incubation, measure the absorbance at 562 nm using a spectrophotometer or plate reader.

6. Create a standard curve using the absorbance values for each standard. Plot the data and perform linear regression analysis.

7. Calculate the protein concentrations of your samples using the standard curve and the absorbance values obtained.

[EasyTools] Drug Dosage Calculator

Drug Dosage Calculator

Enter the following information and click the "Calculate" button to determine the drug volume to administer:

• Drug Concentration (mg/ml): Enter the concentration of the drug solution in milligrams per milliliter (mg/ml). For example, if it's 10 mg of drug in 1 ml of solution, enter 10 mg/ml.
• Mouse Weight (g): Enter the weight of the mouse in grams (g) that will receive the drug.
• Desired Dose (mg/kg): Enter the desired dose in milligrams per kilogram (mg/kg) of the mouse's body weight. For example, if you want to administer 20 mg of drug per kilogram of mouse body weight, enter 20 mg/kg.

Drug Concentration (mg/ml):

Mouse Weight (g):

Desired Dose (mg/kg):

Result: 0

[EasyTools] Solution Calculator

Solution Calculator

Enter the weight of the chemical, molecular weight, and the desired concentration to calculate the required solution volume.

Desired Concentration: M mM uM
Sample Amount (mg):
Molecular Weight (MW):

[EasyTools] Solution Dilution Calculator

Solution Dilution Calculator

The solution dilution calculator calculates the volume of stock concentrate to add to achieve a specified volume and concentration. It uses the formula M1V1 = M2V2, where "1" represents the concentrated conditions (i.e., stock solution molarity and volume), and "2" represents the diluted conditions (i.e., desired volume and molarity).

Stock concentration: M mM uM nM
Desired final volume: L mL uL
Desired concentration: M mM uM nM

[EasyTools] RNA concentration

RNA and DW Calculator

This simple calculator allows users to input RNA concentrations in ug/ul, and it calculates the amount of RNA needed for cDNA synthesis as well as the required volume of DW (distilled water). It can calculate the volume of reagents needed for cDNA synthesis and enables the download of both the entered RNA concentrations and the calculated RNA values.

RNA Measurements (ug/ul):

RNA Amount for cDNA Synthesis (ug):

RNA Amount for cDNA (ug):

Results:

Sample	RNA Amount for cDNA (ul)	DW Volume (ul)

Complementary DNA Sequence Generator

Description: The "Complementary DNA Sequence Generator" is a web-based tool that allows users to generate the complementary DNA (cDNA) sequence of a given DNA sequence. It is a simple and user-friendly utility designed to assist in molecular biology and genetics research. Users can follow these steps to utilize the tool:

Usage Instructions:

1. Enter your DNA sequence in the input field above.
2. Click the "Generate" button to create the complementary sequence.
3. The complementary sequence will be displayed below.

Enter DNA Sequence:

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Complementary Sequence:

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