Pronetbio's Yeast Colony PCR Kit offers an efficient solution for amplifying target genes directly from yeast colonies without DNA extraction. It includes a Yeast lysis buffer and 2× Yeast PCR mix for easy amplification and TA cloning. Fast, reliable, and stable, this kit simplifies your research with minimal handling. Suitable for various yeast species. Available in 40T, 80T, and 320T sizes. Store at -20°C.

INSTRUCTION & COA：

Consult Now

Product Description

Product Overview
The Yeast Colony PCR Kit offers a reliable and efficient solution for amplifying recombinant plasmid target genes from yeast colonies. This kit significantly reduces time and effort in your experiments. It contains a green dye for easy visualization of PCR products in gel electrophoresis, allowing researchers to directly analyze amplification results. The kit includes Yeast lysis buffer for breaking down the yeast cell wall, and 2× Yeast PCR mix that contains PCR buffer, dNTPs, MgCl₂, and hot-start Taq polymerase. Simply add primers and yeast colony template for amplification.
CAT: RY8001
Specifications: 40T / 80T / 320T
Storage Conditions: Store at -20°C, shelf life: 1 year
Product Components:
Components
RY8001-01
RY8001-02
RY8001-03
2×Yeast PCR mix
1 ml
1 ml*2
1 ml*8
Yeast lysis buffer
200 μl
400 μl
1.6 ml
User manual
1 copy
Key Features and Benefits
Fast and Efficient Amplification: Directly amplify target genes from yeast colonies without the need for DNA extraction.
Convenient and Minimal Handling: The 2× Yeast PCR mix simplifies the process, reducing pipetting errors and increasing stability.
High Stability: The 2× Yeast PCR mix remains stable even after repeated freeze-thaw cycles.
Compatible with TA Cloning: PCR products contain a 3' A-tail, suitable for direct TA cloning.
Green Dye for Easy Visualization: The included dye enables easy tracking of PCR products through electrophoresis.
Experimental Protocol
1. Prepare Yeast Cell Lysate:
Add 3-3.5 μl of Yeast lysis buffer into a PCR tube. Using a sterile pipette tip or toothpick, add a small amount of yeast colony to the PCR tube.
2. Incubate at High Temperature:
Mix thoroughly and incubate the tube at 98°C for 10-20 minutes to lyse the yeast cells.
3. Set Up the PCR Reaction:
In a new sterile PCR tube, prepare the following reaction mixture:
2×Yeast PCR mix
25 μL
Primer F*（10 μM） 2 μL
Primer R*（10 μM） 2 μL
Template*
1 μL
ddH₂O
up to 50 μL
* Mix briefly and centrifuge to collect the contents from step 2. Transfer 1 μL of the mix to the PCR reaction as the template.
PCR Conditions:
Place the reaction solution tube into a PCR instrument and run the following program:
Temperature
Time
Cycles
95 ℃ (Initial denaturation)
3 min
1
95 ℃
15 sec

30-32
Tm*
15 sec
72 ℃
15 sec/kb
72 ℃ (Final extension)
5 min
1
4 ℃
∞
1
* Adjust the annealing temperature according to the Tm value of the primer. Typically set the annealing temperature 5°C lower than primer Tm.
Case Studies
Yeast Strain Case
PCR amplification was successfully performed directly from SD/-Leu-Trp-His-Ade media yeast colonies, demonstrating the ease of use of the Yeast Colony PCR Kit for routine genetic analysis.
Bacillus subtilis Case
Our kit was also tested on Bacillus subtilis, a Gram-positive bacterium, showing that the Yeast Colony PCR Kit can be used for rapid amplification without the need for DNA extraction.
Biocontrol Fungi Case
PCR amplification using ITS primers yielded excellent results, confirming the Yeast Colony PCR Kit's versatility in amplifying fungal DNA for research in biocontrol and microbial diversity studies.
FAQs
1. How does the Yeast Colony PCR Kit work?
The Yeast Colony PCR Kit is designed to simplify the amplification of recombinant genes directly from yeast colonies. It includes a yeast lysis buffer to break open yeast cells and a ready-to-use PCR mix that contains all necessary reagents.
2. What is the storage condition for the Yeast Colony PCR Kit?
Store the Yeast Colony PCR Kit at -20°C. The kit is stable for 1 year when stored properly.
3. Can this kit be used for other yeast species?
Yes, the Yeast Colony PCR Kit is compatible with various yeast species. However, PCR conditions such as annealing temperature may need to be optimized depending on the yeast strain and primer sequences.
4. What primers are compatible with the Yeast Colony PCR Kit?
The Yeast Colony PCR Kit can be used with most standard primers for yeast genomic DNA amplification. For best results, optimize the annealing temperature based on the Tm of your primers.
Ready to Advance Your Research?
Contact us today for a detailed consultation and product quotation. When inquiring about this product, please reference Product Code: RY8001.
Disclaimer: All products are for research purposes only. Not for use in diagnostic or therapeutic applications.