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We can simply add this code to show the barcode in the email template:

{% if line.variant.barcode != blank %}
   <span class="order-list__item-variant">Barcode: {{ line.variant.barcode }}</span>
{% endif %}

Under:

{% for line in subtotal_line_items %}

So, It should look like:

{% for line in subtotal_line_items %}
    <!-- Some Code -->
    
    {% if line.variant.barcode != blank %}
        <span class="order-list__item-variant">Barcode: {{ line.variant.barcode }}</span>
    {% endif %}

    <!-- Some Code -->
{% endfor %}

Reference: https://help.shopify.com/en/manual/fulfillment/setup/notifications/email-variables#line-item

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