[EasyTools] PCR Mixture Calculator

Easily calculate the required volumes of DW, Master Mix, and Primers for your PCR experiments. This tool automatically adjusts for pipetting errors based on sample count.

Sample Count (N):

Repeat Count (n):

Select Primer Concentration: 10 pmol stock 100 pmol stock

Calculation Result:

Note: 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 may be adjusted based on objectives and sample types.

* Calculation base: PCR mixture 19 μl + cDNA 1 μl (Total 20 μl reaction)

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 Calculator (BCA Assay)

This tool simplifies Western Blot sample preparation. Input your BCA assay results to automatically calculate protein concentrations, lysate volumes, and required lysis buffer to normalize your samples.

How to Use

1. Enter the raw concentration (ug/mL) for each sample.
2. Input the Dilution Factor used in the assay and the desired Final Volume for normalization.
3. Click Calculate. The tool will highlight the sample with the lowest concentration (limiting factor).

Sample Concentrations (ug/mL)

Sample 1

Sample 2

Sample 3

Sample 4

Sample 5

Sample 6

Sample 7

Sample 8

Sample 9

Sample 10

Common Dilution Factor

Final Sample Volume (uL)

Required 4x Loading Buffer: 0 uL

Normalization Table

Sample	Raw Conc. (ug/mL)	Protein (ug/uL)	Lysate (uL)	Lysis Buffer (uL)

Required Volume for Gel Loading

* Values in parentheses indicate the total volume including 4x Loading Buffer (1.25x).

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

[EasyTools] RNA & cDNA Synthesis Calculator

How to Use:
1. Set your Experimental Condition and Target RNA Amount.
2. Add samples and input concentrations (µg/µL).
3. Click Calculate to get RNA/DW volumes and Master Mix.
4. Negative DW values will be highlighted in red.

Experimental Condition:

Target RNA Amount (µg):

Input RNA Concentration (µg/µL)

Calculation Results

Sample ID	Measured (µg/µL)	RNA Vol (µL)	DW Vol (µL)

[EasyTools] Complementary DNA Sequence Generator

Reverse Complement DNA Generator

This web-based tool generates the Reverse Complement sequence of a given DNA strand. Essential for primer design, probe generation, and sequence analysis in molecular biology. It converts 5'-3' sequences into their anti-parallel complementary strands (5'-3').

How to Use:

1. Paste your DNA sequence (e.g., ATGC...) into the input box.
2. Numbers and whitespace will be automatically removed.
3. Click Generate to see the result.

Input Sequence (5' → 3'):

Length: 0 bp

Output Sequence (5' → 3'):