Cloning heterozygous genes or organisms involves specific strategies to address the challenges posed by the presence of two different alleles at a locus. Here are some approaches based on recent studies:

1. **PCR Amplification and Cloning**

* For heterozygous loci, PCR amplification of the target gene followed by cloning into a plasmid can separate the two alleles. This method avoids ambiguities that arise from directly sequencing heterozygous PCR products. For example, full-length HLA genes have been successfully cloned using this approach.

2. **Sequencing-Based Mapping (SBM)**

In plants, a sequencing-based mapping method has been developed to identify and clone heterozygous mutations. This involves whole-genome sequencing of pooled populations to screen markers, narrowing down candidate regions, and verifying mutations in individual samples. This method is effective for heterozygous mutants in species like Arabidopsis.

3. **Targeted DNA Capture and Sequencing**

For organisms like *Drosophila melanogaster*, targeted DNA capture combined with next-generation sequencing has been used to identify and clone heterozygous mutations. This method significantly reduces the time and cost compared to traditional approaches.

4. **Mitotic Recombination for Homozygous Clones**

Techniques like FLP-FRT recombination in *Drosophila* allow the generation of homozygous clones from heterozygous precursors. This is useful for studying the effects of specific alleles in a controlled genetic background.

5. **Direct Cloning of Amplified Products**

* In cases like beta-thalassemia, the entire gene can be amplified and directly cloned into a plasmid for sequencing. This method ensures accurate identification of mutations, even in compound heterozygous conditions.

These methods provide robust frameworks for cloning and studying heterozygous genes or organisms

The reason you can't find this information is because bacteria plasmids can't be "heterozygous". You can get a mixed population of bacteria with different inserts, or bacteria with more than one type of plasmid (rare, but possible).

You can PCR amplify & clone the WT allele into a plasmid. Do the same for the variant.

To express a gene, you then need to move the region of interest into an expression vector. Now, if your starting organism is eukaryotic (assuming yes?), then you actually need to clone the cDNA, not the gDNA. And you might need to use a vector that is optimized for rare codons.

Please talk with your advisor. I think you need to plan out this project in a lot more detail before you start pipetting anything.